Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Download Author Index
Document related concepts
no text concepts found
Transcript
2014년 대한면역학회 추계학술대회 The 2014 Fall Conference of the Korean Association of Immunologists The Korean Association of Immunologists Host The 2014 Fall Conference of the Korean Association of Immunologists + General Information Conference Venue Sejong University Convention Center in Seoul ·············································· Gwanggaeto building B2F Room Information Plenary Lecture (I, II) ···························································································· B2F Hall A+B Symposium (I, III, V, VII) ······················································································ B2F Hall A Symposium (II, IV, VI, VIII) ·················································································· B2F Hall B Special Lecture (I, III) ···························································································· B2F Hall A Special Lecture (II, IV) ·························································································· B2F Hall B Sponsorship Exhibition & Poster Session ······················································· B2F Hall C Preview ····················································································································· B2F Preview desk i The 2014 Fall Conference of the Korean Association of Immunologists Awards Information KAI 2014 Awards will be presented on Nov. 7, 2014 at 17:30. + Excellent Paper Award SCI citation area 【 RA 】 1. Immune Network 2012 Apr; 12(02) 41-47, Glia as a Link between Neuroinflammation and Neuropathic Pain. Kyoungho Suk, Ph.D. (Kyungpook National University) 【 OA 】 1. Immune Network 2012 Feb; 12(01) 18-26, The Analysis of Vitamin C Concentration in Organs of Gulo−/− Mice Upon Vitamin C Withdrawal. Wang Jae Lee, M.D., Ph.D. (Seoul National University) 2. Immune Network 2012 Feb; 12(01) 8-17, Baculovirus-based Vaccine Displaying Respiratory Syncytial Virus Glycoprotein Induces Protective Immunity against RSV Infection without Vaccine-Enhanced Disease. Jun Chang, Ph.D. (Ewha Womans University) PudMed retrieval area 【 RA 】 1. Immune Network 2014 Feb; 14(01) 21-29, Insights into the Role of Follicular Helper T Cells in Autoimmunity. Je-Min Choi, Ph.D. (Hanyang University) 【 OA 】 1. Immune Network 2013 Aug; 13(04) 123-132, Metformin Down-regulates TNF-∝Secretion via Suppression of Scavenger Receptors in Macrophages. Kyungjae Kim, Ph.D. (Sahmyook University) 2. Immune Network 2013 Aug; 13(04) 133-140, Mesenchymal Stem Cell Lines Isolated by Different Isolation Methods Show Variations in the Regulation of Graft-versus-host Disease. Myung-Shin Jeon, Ph.D. (Inha University) + Best Presentation Award The Korean Association of Immunologists grants award for outstanding posters at the conference. There are 223 abstracts submitted to the poster session of this year’s conference. The Scientific Committee wishes to present awards to all the post presenters, who made efforts in presenting excellent works, but it is a unfortunate that only 20 poster presenters should be selected and awarded through the evaluation of the scientific committee in accordance with the regulation. Posters should be posted up on the board at Convention Hall C between 08:00 a.m. to 09:00 a.m. before the first session of the conference begins. Also, presenters are required to attend and present their works for the evaluation of the Scientific Committee. For presentation schedules, refer to the following information: Poster Session I - Presentation Schedule: 13:20-14:30, November 6 (Thu) - Posters to be Presented: P-001∼P-110, P-203 Poster Session II - Presentation Schedule: 13:20-14:30, November 7 (Fri) - Posters to be Presented: P-111∼P-202, P-204∼P-223 ii The 2014 Fall Conference of the Korean Association of Immunologists Program at-a-Glance Day 1 (Nov. 6, Thu.) 08:00 ~ 08:50 Registration 08:50 ~ 09:00 Opening Address 09:00 ~ 09:50 Plenary Lecture I Peter Cresswell (Yale University, USA) 09:50 ~ 10:20 Coffee Break 10:20 ~ 12:20 Block Symposium I Innate Immunity, Infection and Inflammation Block Symposium II T Cell Development and Function 12:20 ~ 13:20 Luncheon Symposium I Luncheon Symposium II 13:20 ~ 14:30 14:30 ~ 16:30 Poster Session I Block Symposium III Dendritic Cell and Immune Regulation 16:30 ~ 16:50 Block Symposium IV Hot Topics in Transplantation Immunology Coffee Break 16:50 ~ 17:30 Special Lecture I Brian Kelsall (National Institutes of Health, USA) Special Lecture II Shimon Sakaguchi (Osaka University, Japan) 17:30 ~ 18:00 Oral Presentation I Oral Presentation II 18:10 ~ 20:00 Reception iii Day 2 (Nov. 7, Fri.) 08:00 ~ 09:00 Registration 09:00 ~ 09:50 Plenary Lecture II Linda Sherman (The Scripps Research Institute, USA) 09:50 ~ 10:20 Coffee Break 10:20 ~ 12:20 Block Symposium V Autophagy and Regulation of Innate Immunity Block Symposium VI Mucosal Immunology and Vaccine 12:20 ~ 13:20 Luncheon Symposium III Luncheon Symposium IV 13:20 ~ 14:30 14:30 ~ 16:30 Poster Session II Block Symposium VII Clinical Immunology Block Symposium VIII KAI-AKIA Joint Symposium 16:30 ~ 16:50 16:50 ~ 17:30 17:30 ~ 18:00 Coffee Break Special Lecture III Philip D. Hodgkin (Walter and Eliza Hall Institute, Australia) Special Lecture IV Francis R. Carbone (University of Melbourne, Australia) General Assembly, Prize and Closing iv The 2014 Fall Conference of the Korean Association of Immunologists Scientific Program Day 1 (Nov. 6, Thu.) Convention Hall A 08:00-08:50 Registration 08:50-09:00 Opening Address Plenary Lecture I Chair: Chang-Yuil Kang (Seoul National University, Korea) 09:00-09:50 Molecular Mechanisms of MHC Class I-Restricted Antigen Processing Peter Cresswell (Yale University, USA) 09:50-10:20 Coffee Break Block Symposium I: Innate Immunity, Infection and Inflammation Chair: Kee-Jong Hong (Korea National Institute of Health, Korea), Man-Ki Song (International Vaccine Institute, Korea) 10:20-10:50 IL-1 Family Cytokines and Monocytes/Macrophages in the Regulation of Inflammation Diana Boraschi (National Research Council, Italy) 10:50-11:20 New Mechanism of Action and Potential Biomarkers for Vaccine Adjuvant Ken J. Ishii (Osaka University, Japan) 11:20-11:50 Non-Invasive Imaging of Infection, Inflammation and Immune Responses Hyewon Youn (Seoul National University, Korea) 11:50-12:20 Strategies to Tackle Influenza-Invasion of IFN Antagonist Ji-Young Min (Institut of Pasteur, Korea) Luncheon Symposium I 12:20-13:20 Next Generation Technologies for Immuno/Cellular Study and Clinical Application Hak-Jun Ahn (Miltenyi Biotec Korea) Block Symposium III: Dendritic Cell and Immune Regulation Chair: Cheol-Heui Yun (Seoul National University, Korea), Jae-Hoon Choi (Hanyang University, Korea) 14:30-15:05 Common Dendritic Cell Progenitor (CDP), a Novel Source of Dendritic Cells Toshiaki Ohteki (Tokyo Medical and Dental University, Japan) 15:05-15:40 Dendritic Cell and Macrophage Ontogeny Florent Ginhoux (Singapore Immunology Network, Singapore) v 15:40-16:05 Intestinal Aldh1a2+CD103+CD8α+DCs Preferentially Induce Th1cells but Not Tregs Myoung Ho Jang (Academy of Immunology and Microbiology [AIM], Institute for Basic Science [IBS], Pohang, Korea) 16:05-16:30 CTCF Controls Homeostatic Maintenance and Migration of Langerhans Cells in the Skin Hyoung-Pyo Kim (Yonsei University, Korea) 16:30-16:50 Coffee Break Special Lecture I 16:50-17:30 Chair: Pyeung Hyeun Kim (Kangwon National University, Korea) Phenyotype and Function of Mononuclear Phagocytes in the Intestine Brian Kelsall (National Institutes of Health, USA) Oral Presentation I Chair: Seung-Hyo Lee (KAIST, Korea) 17:30-17:40 Functional Heterogeneity of Influenza Virus-specific CD8+ T Cells Jihye Kim (Korea Advanced Institute of Science and Technology) 17:40-17:50 Deficiency of Melanoma Differentiation-associated Protein 5 (MDA5) Results in Exacerbated Chronic Postviral Lung Inflammation Won-keun Kim (University of Pennsylvania) 18:10-20:00 Reception Convention Hall B 08:00-08:50 Registration 08:50-09:00 Opening Address Plenary Lecture I Chair: Chang-Yuil Kang (Seoul National University, Korea) 09:00-09:50 Molecular Mechanisms of MHC Class I-Restricted Antigen Processing Peter Cresswell (Yale University, USA) 09:50-10:20 Coffee Break Block Symposium II: T Cell Development and Function Chair: I-Cheng Ho (Brigham and Women's Hospital, USA), Eun Sook Hwang (Ewha Womans University, Korea) 10:20-11:00 NFAT and PNT Domain-Dependent Regulation of Cytokine Genes by Ets1 I-Cheng Ho (Brigham and Women's Hospital, USA) 11:00-11:30 GATA3, Old-Timer and ZNF131, Newcomer in T Cell Development and Function Shoichiro Miyatake (Tokyo Metropolitan Institute of Medical Science, Japan) 11:30-12:00 Dynamic TGF-β Signaling Networks in T Cell Development and Function Mizuko Mamura (Tokyo Medical University, Japan) vi 12:00-12:20 Molecular Mechanisms Underlying Thymic Involution Eun Sook Hwang (Ewha Womans University, Korea) Luncheon Symposium II 12:20-13:20 Genexine & Technology Won-Seok Lee (Genexine), Dong-Hoon Choi (Genexine) Block Symposium IV: Hot Topics in Transplantation Immunology Chair: Curie Ahn (Seoul National University, Korea), Hong-Rae Choi (Ulsan University Hospital, Korea) 14:30-15:00 Role of Innate Immunity in Ischemia-Reperfusion Injury Byungsuk Kwon (University of Ulsan, Korea) 15:00-15:30 Regulatory T Cell Therapy in Transplantation Kyung-Mi Lee (Korea University, Korea) 15:30-16:00 Mechanisms of Antibody-Mediated Rejection Accommodation in ABO Incompatible Takaaki Kobayashi (Nagoya University, Japan) 16:00-16:30 Immune Monitoring of ABO Incompatible Living Donor Liver Transplant Nayoung Kim (ASAN Medical Center, Korea) 16:30-16:50 Coffee Break Special Lecture II 16:50-17:30 Regulatory T Cells: Development, Maintenance, and Function in Normal and Disease State Shimon Sakaguchi (Osaka University, Japan) Oral Presentation II 17:30-17:40 Chair: Chong-Kil Lee (Chungbuk National University, Korea) Chair: Youngnim Choi (Seoul National University, Korea) Upregulation of PD-1 on Regulatory T Cells Potentiates Their Suppressive Function during Chronic Viral Infection Hyo Jin Park (Yonsei University) 17:40-17:50 Intravital Imaging of T and B Cell Trafficking Across High Endothelial Venules in Mice Lymph Node Kibaek Choe (KAIST) 18:10-20:00 Reception Convention Hall C 13:20-14:30 Poster Session 1 vii Day 2 (Nov. 7, Fri.) Convention Hall A 08:00-09:00 Registration Plenary Lecture II 09:00-09:50 Chair: Charles D. Surh (IBS, Postech, Korea) Mechanisms of Peripheral Tolerance Linda Sherman (The Scripps Research Institute, USA) 09:50-10:20 Coffee Break Block Symposium V: Autophagy and Regulation of Innate Immunity Chair: Eun-Kyeong Jo (Chungnam National University, Korea), Myung-Shik Lee (Sungkyunkwan University, Korea) 10:20-10:55 Roles of Pattern Recognizing Receptors and Autophagy in Antiviral Immune Response Jae Ung Jung (University of Southern California, USA) 10:55-11:25 Autophagy Insufficiency, Inflammasome and Metabolic Syndrome Myung-Shik Lee (Sungkyunkwan University, Korea) 11:25-11:55 Regulation of Innate Immune Response by Autophagy Tatsuya Saitoh (Osaka University, Japan) 11:55-12:20 Small Heterodimer Partner and Innate Immune Regulation Eun-Kyeong Jo (Chungnam National University, Korea) Luncheon Symposium III 12:20-13:20 재택크의 기본은 세택크와 절세의 방법 소개 사승환 지점장 (신한금융) Block Symposium VII: Clinical Immunology Chair: Wan-Uk Kim (The Catholic University of Korea, Korea), Sang-il Lee (Gyeongsang National University, Korea) 14:30-15:00 Approaching the Gene-Specific Therapy of Autoimmunity Richard Bucala (Yale University, USA) 15:00-15:30 βig-h3 and Its Therapeutic Implications in Rheumatoid Arthritis Young Mo Kang (Kyungpook National University Hospital, Korea) 15:30-16:00 Genetic and Immunologic Aspects of Inflammatory Bowel Diseases Jae Hee Cheon (Yonsei University, Korea) 16:00-16:30 Role of Bacteria-Derived Extracellular Vesicles on the Pathogenesis of Chronic Obstructive Airway Disease Yoon-Keun Kim (Ewha Womans University, Korea) 16:30-16:50 Coffee Break viii Special Lecture III 16:50-17:30 Chair: Wang Jae Lee (Seoul National University, Korea) The Cellular Mechanics of the Immune Response Philip D. Hodgkin (Walter and Eliza Hall Institute, Australia) 17:30-18:00 General Assembly, Prize and Closing Convention Hall B 08:00-09:00 Registration Plenary Lecture II 09:00-09:50 Chair: Charles D. Surh (IBS, Postech, Korea) Mechanisms of Peripheral Tolerance Linda Sherman (The Scripps Research Institute, USA) 09:50-10:20 Coffee Break Block Symposium VI: Mucosal Immunology and Vaccine Chair: Charles D. Surh (IBS, Postech, Korea), Sin-Hyeog Im (IBS, Postech, Korea) 10:20-10:50 Immuno-Modulatory Transcriptional Complexes in Regulatory T Cells Dipayan Rudra (IBS [Institute for Basic Sciene]) 10:50-11:20 A New Therapeutic Approach to Asthma Utilizing the TLR5 Signaling Pathways Joon Haeng Rhee (Chonnam National University, Korea) 11:20-11:50 HVEM: a TNF Family Receptor that Controls Diverse Aspects of Mucosal Immunity Mitchell Kronenberg (La Jolla Institute for Allergy & Immunology, USA) 11:50-12:20 IL-17 Family Cytokines in Mucosal Inflammation and Cancer Chen Dong (University of Texas MD Anderson Cancer Center, USA) Luncheon Symposium IV 12:20-13:20 권정근 본부장 (신한금융) 재택크의 기본은 세택크와 절세의 방법 소개 Block Symposium VIII: KAI-AKIA Joint Symposium Chair: Doo Hyun Chung (Seoul National University, Korea), Eui Cheol Shin (KAIST Institute for the BioCentury, Korea) 14:30-15:00 Multifaceted Roles of the Gut Metabolite Short Chain Fatty Acids in Regulation of Mucosal Immunity Chang Hwan Kim (Purdue University, USA) 15:00-15:30 Intracellular Trafficking of Toll-Like Receptors You-Me Kim (POSTECH, Korea) ix 15:30-16:00 Neutrophils Provide Local Chemokine Cues to Regulate CD8 T Cell Responses to Influenza Minsoo Kim (University of Rochester Medical Center, USA) 16:00-16:30 Quantitative and Qualitative Changes in Regulatory T Cell Population during Acute Viral Hepatitis Eui-Cheol Shin (KAIST, Korea) 16:30-16:50 Coffee Break Special Lecture IV 16:50-17:30 Chair: Young Chul Sung (Postech, Korea) Generation and Function of Tissue-Resident Memory T Cells Francis R. Carbone (University of Melbourne, Australia) 17:30-18:00 General Assembly, Prize and Closing Convention Hall C 13:20-14:30 Poster Session 1, 2 x The 2014 Fall Conference of the Korean Association of Immunologists Poster Presentation Antigen Processing and Presentation (P1-4) Allergy, Hypersensitivity and Autoimmunity (P5-24) Immune Cell Development, Differentiation and Function (P25-49) Immune Mediators, Their Receptors and Signaling (P50-60) Immune Response Regulation and Tolerance (P61-94) Innate Immune Response, Infection and Inflammation (P95-151) Mucosal and Regional Immunity (P152-169) Tumor and Transplantation Immunology (P170-192) Vaccines and Immune Therapeutics (P193-210) Technical Innovations and Others (P211-220) Late Breaking Abstracts (P221-223) xi The 2014 Fall Conference of the Korean Association of Immunologists CONTENTS • Layout of the Venue ··································································································· i • Awards Information ···································································································· ii • Program at-a-Glance ································································································ iii • Scientific Program ····································································································· v • Plenary Lecture ········································································································· 1 • Special Lecture ·········································································································· 7 • Block Symposium ··································································································· 17 • Oral Presentation ···································································································· 51 • Poster Presentation ································································································ 57 • Luncheon Symposium ·························································································· 115 • Author Index ··········································································································· 119 xii The 2014 Fall Conference of the Korean Association of Immunologists Nov. 6(Thu) ~ 7(Fri), 2014 Seoul, Korea Plenary Lecture BIOGRAPHY Peter Cresswell Peter Cresswell is the Eugene Higgins Professor of Immunobiology and Professor of Cell Biology at Yale University School of Medicine. He is also an investigator of the Howard Hughes Medical Institute. He received his B.S. degree in chemistry and M.S. degree in microbiology from the University of Newcastle Upon Tyne, U.K., and his Ph.D. degree in biochemistry and immunology from London University. He is a Fellow of the Royal Society, U.K., a member of the National Academy of Sciences, the American Academy of Arts and Sciences and the Institute of Medicine. Dr. Cresswell’s research focuses on the functions of Major Histocompatibility Complex (MHC) molecules and CD1 molecules, which, respectively, bind peptides derived from protein antigens or lipids, forming complexes recognized by T lymphocytes during immune responses. The mechanisms governing ligand association are collectively known as antigen processing. Additional major interests are in the actions of a variety of proteins induced by interferons that play a role in immunity to infection. Plenary Lecture [PL I] Molecular Aspects of MHC Class I-Restricted Antigen Processing Peter Cresswell Eugene Higgins Professor of Immunobiology, Investigator, Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, CT, USA The recognition of virus-infected cells or tumor cells by CD8-positive T lymphocytes relies on the formation of complexes of MHC class I molecules with virus-derived or tumor specific peptides, respectively. The peptides are generated in the cytosol and are translocated into the endoplasmic reticulum (ER), where peptide binding occurs, by the dimeric ATP-dependent Transporter associated with Antigen Processing (TAP). TAP is a component of the multi-subunit Peptide Loading Complex (PLC). The PLC also incorporates the glycoprotein tapasin, a product of an MHC-linked gene, which physically links MHC class I-β2-microglobulin dimers to the PLC. The preferential association of MHC class I molecules with high affinity peptides is mediated by tapasin and is facilitated by an adaptation of the calnexin/calreticulin quality control cycle that normally facilitates the proper folding of a variety of glycoproteins in the ER prior to transport. The lectin chaperone calreticulin and the thiol oxidoreductase ERp57, two components of this folding pathway, are also associated with the PLC and play a critical role in the peptide binding process. The mechanism of action of the PLC and the nature of the quality control mechanisms underlying the formation of MHC class I-peptide complexes will be discussed. Keywords: Antigen presentation, HLA, Protein folding 4 The 2014 Fall Conference of the Korean Association of Immunologists BIOGRAPHY Linda Sherman Dr. Linda Sherman did her Ph.D. in Biology at M.I.T. Following postdoctoral research at Harvard Medical School she took a position at The Scripps Research Institute in La Jolla, California, where her laboratory has been doing basic research in immunology for 35 years. Her research interests have included transplantation, cancer immunotherapy, and autoimmunity. The focus of her laboratory has been to study the underlying causes of autoimmune diseases, such as type 1 diabetes, with the goal of prevention; and also to learn how to harness the self-destructive capability of the immune system for the destruction of tumor cells. Dr. Sherman has trained over 40 postdoctoral fellows and graduate students and published over 110 manuscripts. She has served on numerous advisory boards for government and non-profit institutions, as well as several biotech companies. She was recently elected president of the American Association of Immunologists. Plenary Lecture [PL II] Mechanisms of Peripheral Tolerance Linda Sherman, Grégory Verdeil, Trevor Smith, Christian Maine, Xiaotian Lin Department of Immunology and Microbial Sciences. The Scripps Research Institute, La Jolla, CA, USA Peripheral tolerance prevents autoimmune attack by self-reactive T cells that escape thymic deletion. Tolerance mechanisms intrinsic to the T cell include T cell inactivation (anergy), deletion and ‘tuning’ of TCR signal transduction. The distinguishing molecular features that determinethe difference between deletion and anergy are not known. Using an in vivo tolerance model in which anergy and deletion of CD8 T lymphocytes is determined by antigen dose or strength of signal,we identified features unique to each mechanism. As compared with cells undergoing deletion, anergic cells exhibited increased levels of CD122 and demonstrated responsiveness to IL-15, suggesting its serves as the survival cytokine for anergic cells. Anergic CD8 T cells also exhibited increased expression of negative co-stimulatory molecules characteristic of expressed of exhausted CD8 T cells, including PD-1, Tim-3 and NKG2A. Cells undergoing deletion exhibited increased expression of the pro-apoptotic molecule BIM, and reduced levels of expression of numerous proteins involved in cholesterol metabolism. Recently we have also examined the role of the protein tyrosine phosphatase, PTPN22 in maintaining T cell tolerance by ‘tuning’ down TCR signaling. It appears to play a particularly important role in reducing the strength of TCR signal in mouse and human memory T cells so as to prevent T cell activation in response to self-antigens. 6 The 2014 Fall Conference of the Korean Association of Immunologists The 2014 Fall Conference of the Korean Association of Immunologists Nov. 6(Thu) ~ 7(Fri), 2014 Seoul, Korea Special Lecture BIOGRAPHY Brian Kelsall Dr. Brian Kelsall received his B.A. in human biology from Stanford University in 1982. In 1986, he earned his M.D. from Case Western Reserve University School of Medicine. He did postdoctoral training in internal medicine at The New York Hospital-Cornell Medical Center from 1986 to 1989 and in infectious diseases at the University of Virginia Medical Center from 1989 to 1992. In 1992, Dr. Kelsall went to the National Institutes of Health, completed fellowship training in mucosal immunology in 1996, and became a senior investigator in 2003. His research focuses on the regulation of immune responses in the intestine, in particular the role that unique intestinal dendritic cell and macrophage populations play in the induction of immunity to intestinal viral pathogens and mucosal vaccines and in the pathogenesis of inflammatory bowel disease. Special Lecture [SL I] Phenotype and Function of Mononuclear Phagocytes in the Intestine Alessandra Filardy, Abhi Kole, Americ Rivollier, Kazuya Kitamura, Jianping He, Brian Kelsall Mucosal Immunobiology Section, Laboratory of Molecular Immunology, NIAID, NIH, Bethesda, MD, USA We, and others have previously demonstrated the presence of different populations of mononuclear phagocytes in the intestine, including the small and large intestinal lamina propria, the Peyer’s patches, mesenteric lymph nodes (MLNs), and the solitary intestinal lymphoid tissues (SILT). This lecture will focus on cell populations in the colon, and discuss novel aspects of their function, including the regulation of inflammatory and regulatory cytokine genes, with a focus on TNFα, IL-6, IL-1βand IL-10. Specifically, we found a role for IFN-1 in controlling IL-10 and IL-1RA production by intestinal macrophages, that affects DC activation and T cell priming in the MLNs with consequences for regulatory and effector T cell differentiation. In addition, we have identified that pro-inflammatory gene expression by intestinal macrophages in the steady-state is controlled largely at a post-transcriptional level that involves enhanced protein degradation. Details of these studies will be discussed. 10 The 2014 Fall Conference of the Korean Association of Immunologists BIOGRAPHY Shimon Sakaguchi Shimon Sakaguchi is a Distinguished Professor at the World Premier International Research Initiative (WPI)-Immunology Frontier Research Center (IFReC) at Osaka University, Japan. He is an immunologist recognized for his work on the control of immune responses. He is known particularly for his discovery of regulatory T cells, an indispensable constituent of the immune system for the maintenance of immune self-tolerance and homeostasis. Sakaguchi was born in Japan in 1951, obtained an M.D. in 1976 and a Ph.D. in 1982 from Kyoto University, Japan, where he was trained as a pathologist and immunologist. After performing postdoctoral studies at Johns Hopkins University and Stanford University as a Lucille P. Markey Scholar, he served as an Assistant Professor in the Department of Immunology at the Scripps Research Institute. He returned to Japan in 1991 and continued his immunology research at RIKEN Institute as an Investigator of the Japan Science and Technology Agency and subsequently as the Head of the Department of Immunopathology at Tokyo Metropolitan Institute of Gerontology, Tokyo. From 1998 to2011, he was a Professor and the Chairman of the Department of Experimental Pathology, Institute for Frontier Medical Sciences Kyoto University and served as the Director of the Institute for several years. In 2011, his lab moved to Osaka University and he assumed the current position. Research Interests: Sakaguchi studies the molecular and cellular mechanisms of immunological tolerance and immune regulation. He has shown that a population of immunosuppressive T-lymphocytes, designated regulatory T cells, are present in the immune system and its deficiency or dysfunction is causative of a variety of immunological disorders including autoimmune diseases. He has investigated the molecular basis of regulatory T cell development and function. In addition, Sakaguchi and his laboratory have demonstrated that numerical expansion of regulatory T cells or strengthening of their suppressive activity is capable of preventing and treating autoimmune diseases and also establishing stable tolerance to transplanted organs, while their reduction in number or suppressive activity is able to provoke effective immunity against cancer. Sakaguchi is currently investigating how regulatory T cells can be targeted in humans to control a variety of physiological and pathological immune responses in clinical settings. Special Lecture [SL II] Regulatory T Cells: Development, Maintenance, and Function in Normal and Disease States Shimon Sakaguchi Immunology Frontier Research Center, Osaka University, Japan Naturally occurring regulatory T (Treg) cells, which express the transcription factor Foxp3, are indispensable for immunological unresponsiveness to self-constituents, i.e., self-tolerance, and immune homeostasis. They however hamper effective tumor immunity because many tumor-associated antigens are antigenically normal self-constituents. It is therefore a key issue to determine how effective tumor immunity can be evoked without deleterious autoimmunity. By addressing the issue, we have characterized subpopulations of human Foxp3+ T cells, which included suppressive and non-suppressive populations, as well as cell fates of responder T cells controlled by Treg cells. Our study has shown that Foxp3+ cells can be separated into at least three functionally distinct subpopulations, including (1) highly proliferative and differentiated Foxp3-high population (designated effector Treg cells) with stable Treg-specific epigenome alterations, the majority of which will die by apoptosis after suppression, (2) naïve Foxp3-low population (naïve Treg cells), which is in a resting state and upon TCR stimulation differentiates into effector Treg cells, and (3) Foxp3-low non-suppressive population unstable in Treg-specific epigenome changes and capable of secreting proinflammatory cytokines. By searching for specific markers for each population, we will show that each population exhibits distinct behavior in different disease states, and that depletion of effector Treg cells alone, by preserving naïve Treg cells, is sufficient for evoking strong tumor immunity without eliciting serious autoimmunity. We will discuss how Treg subpopulations can be controlled for augmenting or suppressing a variety of immune responses in clinical settings. 12 The 2014 Fall Conference of the Korean Association of Immunologists BIOGRAPHY Philip D. Hodgkin Phil Hodgkin is an experimental and theoretical immunologist and current head of the Immunology Division at the Walter and Eliza Hall Institute, Melbourne. His primary research interests centre upon the regulation of T and B lymphocytes. His laboratory develops quantitative methods for exploring lymphocyte behaviour including single cell imaging and division-based tracking. His laboratory is developing analytical software tools for simulating the effect of cytokines and genetic changes on lymphocyte growth, survival and differentiation. His goal is to understand the behaviour of lymphocytes and recreate the immune response with computer-based models. Special Lecture [SL III] The Cellular Mechanics of the Immune Response 1,2 1,2 1,2 Philip D. Hodgkin , Andrey Kan , Julia M. Marchingo , Jie H.S. Zhou, 3 1,2 1,2 4 John. F. Markham , Mark R. Dowling , Susanne Heinzel , Ken R. Duffy 1 2 The Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia; The 3 University of Melbourne, Melbourne, Victoria, Australia; Victoria Research Laboratory, National Information and Communications Technology (ICT) Australia, University of Melbourne, Melbourne.; 4 Hamilton Institute, National University of Ireland, Maynooth, Ireland During the adaptive immune response T and B-lymphocytes receive signals from different sources that eventually determine the strength and type of response they follow. Our laboratory has been studying the rules of signal integration and decision-making under different conditions. By combining studies of population changes over time, both in vitro and in vivo, with direct imaging of single cells making decisions under different stimulation combinations a picture of how T and B cells operate as ‘machines’ and respond to regulation is becoming clear. Perhaps surprisingly cellular calculation can be described with quite simple quantitative rules, although there are many ‘variations’ and complex outcomes possible. In the most fundamental response B and T cells simply count through a series of divisions, stop, return to quiescence and die. Here the number of times T cells divide is perhaps the most important variable and is highly influenced by stimulation strength and the unique combination of costimulatory signals. Furthermore we report a role for stochastic variation of timed processes to divide, die and differentiate to manipulate the strength of the population response and to regulate the proportion of cells allocated to different tasks. While perhaps impossible to predict the infinite variety of paths taken by single cells and families, knowledge of the underlying stochastic processes, combined with rules of integration acquired by single cell tracking and analysis enables accurate modelling of the population response over time. 14 The 2014 Fall Conference of the Korean Association of Immunologists BIOGRAPHY Francis R. Carbone Dr. Francis Carbone works in the area of T cell recognition and antigen presentation. He defined the interconnection between dendritic cells and cross-presentation for tolerance induction and the response against infection, showing that selection between immunity and tolerance is determined by the activation state of the dendritic cells. More recently, he has focused on means of generating effective immunity in peripheral tissues. He has shown that there are distinct populations of memory T cells permanently resident in tissues such as skin and that these cells provide a rapid response at first pathogen encounter. His laboratory is currently determining how these cells are lodged in peripheral tissues and defining the mechanisms responsible for their effectiveness and long-term survival. Special Lecture [SL IV] Generation and Function of Tissue-Resident Memory T Cells Francis R. Carbone, Thomas Gebhardt, Laura Mackay Department of Microbiology and Immunology, The University of Melbourne, Australia Tissue-resident memory T cells (TRM) can be found in a variety of different extra-lymphoid organs. In our experi+ + ments, we found that CD8 CD103 TRM cells develop in the skin from killer lectin-like receptor G1 (KLRG-1)-negative precursors that selectively infiltrate the epithelial layer. These cells alone conferred enhanced protection against virus infections at body surfaces. A combination of epithelial entry in addition to interleukin 15 (IL-15) and transforming growth factor-β (TGF-β) signalling was required for optimal formation of these long-lived memory cells in skin. Importantly, TRM differentiation resulted in the progressive acquisition of a unique transcriptional profile that differed from those expressed by circulating memory cells. Selective expression of a unique set of transcriptional factors dramatically influenced TRM formation and/or survival. Combined, these data provide a molecular framework for the local differentiation of a distinct peripheral memory population that forms a first-line immune defence system in barrier tissues. 16 The 2014 Fall Conference of the Korean Association of Immunologists The 2014 Fall Conference of the Korean Association of Immunologists Nov. 6(Thu) ~ 7(Fri), 2014 Seoul, Korea Block Symposium Partially supported by Seoul National University internal fund. Innate Immunity, Infection and Inflammation Block Symposium I BIOGRAPHY Diana Boraschi Diana Boraschi is an immunologist that built her experience both in academic institutions (Italian National Council for Nuclear Energy, Rome, Italy; National Cancer Institute in Bethesda, MD; Mario Negri Institute in Milan, Italy; University of Michigan Medical School, Ann Arbor, MI; CNR in Pisa) and industrial settings (the vaccine company Sclavo in Siena, Italy; the pharmaceutical company Dompé in L’Aquila, Italy). She is presently Research Director at the Institute of Protein Biochemistry of the Italian National Research Council in Naples. She has served as Director of Fellowships at the Human Frontier Science Program Organization in Strasbourg, France. As external expert evaluator, she has served in the research programmes of the EU Commission (FP5, FP6, FP7), EDCTP, the Singapore National Medical Council, the US National Science Foundation, and governmental initiatives in Egypt, Romania, Sweden, Poland, South Africa, and Italy. She also serves as peer reviewer for numerous scientific journals and is on the editorial board of seven of them. She is author of 153 peer-reviewed research articles in immunology, editor of 17 books, and inventor in eight patents, in addition to numerous monographic and divulging publications. Since many years she is involved in international higher education training activities, with particular emphasis on capacity building actions in Africa in the field of poverty-related diseases and health care systems and delivery. Diana Boraschi studies the mechanisms of innate defence responses, focussing in particular on the role of macrophages and inflammatory cytokines in the effector phase of defence reactions against infections and tumours. Her main interests are the receptors of the IL-1R/TLR family and their cytokine ligands (IL-1 and IL-18). A fragment of IL-1 endowed with immunostimulatory activity is now defined as the “Boraschi loop”. She is currently studying the role of inflammation in the pathogenesis of diseases (from autoimmune syndromes to degenerative diseases such as ALS), with emphasis on abnormalities in the activation of macrophages. Within the study of the initiating mechanisms causing chronic inflammatory and autoimmune diseases, she has addressed the possible impact of engineered nanoparticles, and of their interaction with microbial derived factors, in initiating or modulating pathology-related inflammation. She has initiated the “Immunosafety Focus Group” within the NanoSafety Cluster (an initiative sponsored by the EU Commission), aiming at defining and standardising the immunosafety assessment as central part of nanosafety regulations. [BS I-1] IL-1 Family Cytokines and Monocytes/Macrophages in the Regulation of Inflammation Diana Boraschi1*, Ettore Mosca2, Roberta Alfieri2, Luciano Milanesi2, Paola Italiani1,2 1 2 Institute of Protein Biochemistry, National Research Council, Napoli, Italy, Institute of Biomedical Technologies, National Research Council, Segrate, Milano, Italy The IL-1 family encompasses eleven ligands and ten receptors, which have a key role in different phases of the acute resolving inflammatory defence responses, as well as in the persistent inflammation characteristic of chronic inflammatory diseases. Both inflammatory and an- ti-inflammatory cytokines are present among the ligands of the IL-1 family, including natural inhibitors/antagonists. IL-1 family receptors include ligand-binding activating receptors, non-binding accessory chains, and ligand-binding non-signalling “decoy” receptors, the latter being able to subtract the agonist ligands from interaction with activating receptors. The proper quantitative and temporal modulation of all these molecules underlies the physiological resolving inflammatory defensive responses, while an imbalance may result in the transition to chronic/pathological inflammation. The human innate/inflammatory response in a tissue can be modelled in vitro by culturing human normal monocytes isolated from blood in conditions that resemble the recruitment from blood into the inflamed site, then the encounter with the inflammatory agents and the development of an inflammatory reaction, and eventually the conditions promoting tissue repair and homeostatic regulation upon resolution. In order to simulate + o this sequence of events, CD14 monocytes were exposed sequentially in culture to CCL2 (from 0 to 2 h, 37 C, normoxia), LPS-displaying bacterial o o vesicles (from 2 to 14 h, 39 C, hypoxia), TNF-α (from 3 to 14 h, 39 C, hypoxia), IFN-γ (from 7 to 14 h, 39oC, hypoxia), IL-10 (from 14 to 24 h, 37oC, normoxia), and TGF-β (from 24 to 48 h, 37oC, normoxia). Moreover, by changing the culture conditions, we also reproduced the con+ o ditions of persistent/chronic inflammation, such as in rheumatoid arthritis, by exposing CD14 monocytes in culture to CCL2 (from 0 to 2 h, 37 C, o normoxia), a cocktail of bacterial and viral stimuli (LPS, PDG, poly:IC, CpG, from 2 to 7 h, 39 C, hypoxia) and IFN-γ, ACPA-PA immune complexes, Survivin, M-CSF and GM-CSF (from 7 to 96 h, 39oC, hypoxia). The modulation of the IL-1 family members in monocytes during the different phases of acute and chronic inflammatory reactions was profiled by transcriptome analysis using RNA-Seq. Expression of the genes for six cytokines (IL1B, IL1A, IL1RN, IL18, IL18BP, IL36G) and four receptors (IL1R1, IL1R2, IL1RAP, SIGIRR) are similarly regulated during the early phase of the acute and chronic inflammatory responses, with the peak of expression between 2 and 14 h. The expression of most of these genes returned to the low basal level during the resolution phase in the acute model (except IL18 that was less expressed than in basal conditions), while it was higher than in fresh monocytes in the chronic model from 24 h on. Exceptions are the genes of the two inhibitors IL18BP and SIGIRR. The former significantly increased from 24 to 96 h, while the latter decreased throughout the entire course of the inflammatory response. The GSEA analysis revealed that in both acute and chronic inflammation the expression of all the IL-1 family members is mainly modulated during the first fourteen hours of stimulation, a period in which it is possible to observe the largest variation of expression. Moreover, the analysis of differentially expressed genes between two consecutive time points confirmed that IL-1 family members are modulated during the entire course of the inflammatory reactions, except between 72 and 96 h in the chronic model where they remain practically unchanged. In conclusion, our results confirm the strong involvement of the IL-1 family genes, especially in the early phases of the monocyte inflammatory response, to underline the central role of IL-1 related cytokines and receptors in initiating innate/nflammatory defensive reactions. This work was supported by the EU FP7 project BioCog (GA 602461), by the Cluster project Medintech of the Italian Ministry of University and Research (CTN01_00177), and by the Italy-Korea Bilateral GR project on Nanovaccination and Adjuvants. Keywords: Monocytes, Macropahges, Interleukin-1, Inflammation, Resolution The 2014 Fall Conference of the Korean Association of Immunologists 19 Supported from International Vaccine Institute. Block Symposium I Innate Immunity, Infection and Inflammation BIOGRAPHY Ken J. Ishii Prof. Ken Ishii is currently a Project leader at the Laboratory of Adjuvant Innovation at the National Institute for Biomedical Innovation (NIBIO) as well as a Professor at the Laboratory of Vaccine Science at the Immunology Frontier Research Center (IFREC) of Osaka University in Japan. Prof. Ishii graduated with a M.D and also obtained a Ph.D. from the School of Medicine, Yokohama City University, Kanagawa, Japan. He has worked on basic immunology as well as translational science of vaccines and adjuvants for last 18 years and have published more than 130 peer-reviewed publications cited over 10000 times leading to my h-index 47 (2014). He is further qualified with his years of experience in vaccine research and regulation including 7 years as a Visiting Scientist and IND reviewer at the Office of Vaccine Research and Review (OVRR), Center for Biologics Evaluation and Research (CBER), US Food and Drug Administration (FDA). Based on his career, he conducts basic research on innate immune recognition of adjuvants and try to translate into vaccine markets in Japan. [BS I-2] New Mechanism of Action and Potential Biomarkers for Vaccine Adjuvant Ken J. Ishii1,2 1 Laboratory of Adjuvant Innovation, National Institute of Biomedical Innovation (NIBIO), Laboratory of Vaccine Science, Immunology Frontier Research Center (IFREC), Osaka University, Osaka, Japan 2 The word adjuvant has its origin from the Latin "adjuvare", meaning "to help". It is a general term for substances (factors) which are co-administered with a vaccine with the aim of increasing the effect (immunogenicity) of the vaccine. The research and development of adjuvants has a history of more than 80 years, and their actual mechanism was not immunologically understood for a long time, with a famous sarcastic remark "Immunologist's dirty little secret". Recent advance in Immunolgy; however, allowed the development of adjuvants through an innovative scientific approach, and there is fierce competition worldwide for the development of next-generation adjuvants. I would like to introduce and discuss about several adjuvants with their novel mechanisms, including a small compound as a potent DAMPs inducer to target certain innate immune mechanisms. On the other hand, however, adjuvants range widely in terms of origin and mode of action, and they may be the cause or underlying cause of vaccine toxicity, especially immunotoxicity. I will present our recent work that common particulate adjuvants can cause local but sustained inflammation and allergic responses via novel innate immune mechanisms and cell death. References 1. Kobiyama K et al Nonagonistic Dectin-1 ligand transforms CpG into a multitask nanoparticulate TLR9 agonist. Proc Natl Acad Sci U S A. 2014 111(8):3086-91. 2. Kuroda E, Coban C, Ishii KJ. Particulate adjuvant and innate immunity: past achievements, present findings, and future prospects. Int Rev Immunol. 2013 32(2):209-20. 3. Jounai N, et al. Recognition of damage-associated molecular patterns related to nucleic acids during inflammation and vaccination. Front Cell Infect Microbiol. 2012;2:168. 4. Desmet CJ, Ishii KJ. Nucleic acid sensing at the interface between innate and adaptive immunity in vaccination. Nat Rev Immunol. 2012 12(7):479-91. 5. Marichal T et al. DNA released from dying host cells mediates aluminum adjuvant activity. Nat Med. 2011 17(8):996-1002. 20 The 2014 Fall Conference of the Korean Association of Immunologists Innate Immunity, Infection and Inflammation Block Symposium I BIOGRAPHY Hyewon Youn Hyewon Youn is an associate professor of Cancer Imaging Center at Seoul National University Hospital. She also has an joint appointed with Department of Nuclear Medicine, Seoul National University College of Medicine. She received B.S., M.S. from Seoul National University and Ph.D. from Texas Tech Univ. She worked for Texas Tech Health Sciences Center as a postdoc. She served at SouthWest Cancer Center and Univ. of Kansas Medical Center as a research scientist and an instructor. She joined the Department of Nuclear Medicine in Seoul National University College of Medicine at 2008. The primary goal of her research is to translate the acquired knowledge from basic research into clinic using molecular imaging. Specifically, her current research interests include 1) the elucidation of molecular events that govern the interaction between cells and their microenvironment 2) the tracking of pathogens, stem cells and leukocytes; 3) the evaluation of therapeutic effect including chemo-, radio-, gene-, stem cell, immune- and their combined applications; 4) the development of imaging reporters & theragnostic probes for multimodal imaging. [BS I-3] Non-Invasive Molecular Imaging of Infection, Inflammation and Immune Responses Hyewon Youn Seoul National University, Korea Clinical and preclinical in vivo imaging approaches have been used to study biological responses at the microscopic (intra-vital imaging) and macroscopic (whole-body imaging) level using various labeling method. A series of imaging techniques ranging from non-radiation based techniques such as optical imaging, MRI, and ultrasound to radiation based CT/nuclear imaging can be used for in vivo imaging. These imaging modalities highlight the intrinsic behavior of different cell populations in physiological context. Fluorescent, radioactive or paramagnetic probes can be used in direct labeling protocols to monitor the specific cell population. Reporter genes can also be used for genetic, indirect labeling protocols to track the fate of a given cell subpopulation in vivo. In this talk, I will briefly summarize several methods to visualize pathogens and immune cells at the level of macroscopic whole-body imaging for the study of their physiological function in the context of infection and immunity, and illustrate our recent studies to exploit imaging-derived information. The 2014 Fall Conference of the Korean Association of Immunologists 21 Block Symposium I Innate Immunity, Infection and Inflammation BIOGRAPHY Ji-Young Min Dr. Ji-Young Min received her doctoral degree from the University of Texas at Austin, US and completed her first fellowship from the Institute of Cellular and Molecular Biology at the same University and the second fellowship from the Laboratory of Infectious Diseases (LID), NIAID, NIH, US. After postdoctoral training she served as Staff Scientist of Emerging Respiratory Viruses Section of LID, NIAID, NIH. Dr. Min joined the Institute Pasteur Korea (IPK) as a Senior Investigator in 2011. She is a member of American Society of Microbiology and American Society for Virology. She received NIH Merit Award 2008 and 2009 and Young Influenza Scientist Recognition Award by European Society of Influenza in 2011. Her research themes spans from the biology of the influenza viruses in host immune system and the development of vaccines against pandemic strains of influenza in preclinical and clinical trials. Dr. Min has extensive experience of working with biosafety level 3 selective agents and conducted studies to elucidate mechanism of transmission and adaptation of highly pathogenic avian influenza viruses. At the IPK, Dr. Min’s research is focused on the development of antiviral agents against seasonal and pandemic strains of influenza. [BS I-4] Strategies to Tackle Influenza: A Nexus between Basic Science and Clinical Application Ji-Young Min Respiratory Viruses Research Group, Institut Pasteur Korea Discovery and utilization of small molecule compounds are becoming increasingly important part of basic biological researches. When executed judiciously, screening for and optimization of small molecule hits can provide a powerful tool compound by which the latent biology can be dissected to further our understanding of the biological processes. The NS1 protein of influenza viruses plays a multifunctional role in host-pathogen interaction and has recently been explored as a potential target for antiviral drug development. The dsRNA binding property of NS1 is considered an attractive target for antiviral drugs since it has been implicated in IFN-βantagonism and immune evasion. Virtual docking of 30,000 small molecules chemical libraries against the NS1 dsRNA-binding pocket revealed several candidate scaffolds that may interact with this region and potentially interfere with its function. One class of molecules exhibited antiviral activities against the recombinant influenza A/Puerto Rico/8/1934 NS-GFP H1N1 virus, contemporary seasonal influenza strains A/California/07/2009 H1N1 and A/Perth/16/2009 H3N2, the previous seasonal strain A/Udorn/307/1972 H3N2, as well as the avian influenza A/Aquatic Bird/Korea/CN2-P1L7 H5N2 virus in cell-based assays. Interestingly, treatment of influenza-infected cells with dsRNA binding inhibitor elevated the levels of some cytokines and chemokines compared with infection alone. Taken all together, the NS1 RNA binding domain should be further explored as a potential target for future drug development programs. Presented in this talk are innovative phenomic approaches that are not only helping to shed a new light on the discovery of first-in-class compound with novel molecular mechanism of influenza virus infection, but also providing a great hope of fighting some of the more devastating pandemic in new ways. 22 The 2014 Fall Conference of the Korean Association of Immunologists Hosted by BK21+ Frontier Pharmacy Leaders, Chungbuk National University. T Cell Development and Function Block Symposium II BIOGRAPHY I-Cheng Ho Dr. I-Cheng Ho is an associate professor of Harvard Medical School and a rheumatologist at Brigham and Women’s Hospital. His research interest is in understanding the molecular mechanisms mediating autoimmune diseases, particularly rheumatoid arthritis and lupus. He also sees patient at the Arthritis Center of Brigham and Women’s Hospital at Boston and Brigham and Women’s/Mass General Health Care Center at Foxborough. [BS II-1] NFAT and PNT Domain-Dependent Regulation of Cytokine Genes by Ets1 I-Cheng Ho Brigham and Women's Hospital, USA Ets-1, the prototype of the ETS family of transcription factors, is required for the expression of IL-2 but suppresses the production of Th17 cytokines. It contains a Pointed (PNT) domain, a protein-protein interacting domain, which is highly conserved among several ETS members. However, the molecular mechanism mediating the function of Ets1 and the role of its PNT domain have remained unclear. We have found that Ets-1 physically and functionally interacts with the nuclear factor of activated T-cells (NFAT) and is required for the recruitment of NFAT to the IL-2 promoter. In addition, Ets-1 is located in both the nucleus and cytoplasm of resting Th cells. Nuclear Ets-1 quickly exits the nucleus in response to calcium dependent signals and competes with NFAT proteins for binding to protein components of NRON (non-coding RNA repressor of NFAT) complex, which serves as a cytoplasmic trap for phosphorylated NFAT proteins. This nuclear exit of Ets-1 precedes rapid nuclear entry of NFAT and Ets-1 deficiency results in impaired nuclear entry but not the dephosphorylation of NFAT proteins. We have also discovered that the PNT domain of Ets1 is dispensable for the development of T cells in vivo but is required for Ets1 to suppress the expression of IL-17 by Th17 cells. Thus, Ets1 regulates the expression of cytokines genes by both NFAT and PNT domain-dependent mechanisms; targeting the PNT domain of Ets1 can modulate the function of Th cells without interfering with their development. The 2014 Fall Conference of the Korean Association of Immunologists 23 Block Symposium II T Cell Development and Function BIOGRAPHY Shoichiro Miyatake 【 Education 】 1981 1983 1989 B.S., University of Tokyo, Japan M.S., University of Tokyo, Japan Ph.D., University of Tokyo, Japan 【 Career 】 1987∼1991 1991∼1996 1996∼1997 1997∼2000 2000∼2010 2010∼Present Postdoc, DNAX Research Institute, USA Assistant Professor, Department of Medicine, Chiba University, Japan Lecturer, Department of Medicine, Chiba University, Japan Research fellow, Institute of Medical Science, University of Tokyo Project Leader, Tokyo Metropolitan Institute of Medical Science, Japan Head, Laboratory of Self Defense Gene Regulation, Tokyo Metropolitan Institute of Medical Science, Japan [BS II-2] GATA3, Old-Timer and ZNF131, Newcomer in T Cell Development and Function Tomohiro Iguchi, Kazuhisa Aoki, Shoichiro Miyatake Laboratory of Self Defense Gene Regulation, Tokyo Metropolitan Institute of Medical Science, Japan Transcription factor GATA3 has multiple roles in T cell development and function. It is required for early T cell development and the choice of CD4SP differentiation in thymus. In periphery Th2 subset induction requires GATA3. It also plays some role in CD8 T cell function as well as in Treg function. One of the approaches to elucidate its function is to look for GATA3 interacting molecules. We identified a member of BTB/POZ Zn finger protein family, ZNF131 as one of the GATA3 partner candidates. When znf131 is deleted during DN2/3 stages of T cell development by crossing with Lck-cre mice, DN to DP differentiation accompanied with proliferative expansion was abolished. We found that CDK inhibitor, cdkn1a, was deregulated. When znf131 was deleted in bone marrow by crossing with Mx-cre mice and the cre-mediated deletion was induced by pI-pC injection, the decrease of cellularity of the very early stage of T cell development in thymus was observed. However, T cell precursor populations before homing to thymus were not affected. Those data indicate that up to DP stage znf131 or gata3 deletion showed very similar phenotype, suggesting that GATA3 work together with ZNF131 to regulate a certain group of targets presumably required for proliferation. When znf131 was deleted during DP stage, there is no obvious impairment of the thymic selection and the maturation of T cells in thymus. However the number of peripheral T cells was significantly reduced. After the DP stage, inactivation of gata3 or znf131 revealed different phenotype. The ratio of memory phenotype T cells that retain intact znf131 allele was increased, indicating that homeostasis of T cells was impaired. Furthermore the proliferation upon activation of T cells through TCR signal was severely impaired. We found the deregulation of cdkn1a again. Our hypothesis is that one of the mechanisms that ZNF131 regulates cell proliferation is through its suppression of cdk inhibitor, cdkn1a. I would like to discuss our GATA3KO mice that develop autoimmune dermatitis also. 24 The 2014 Fall Conference of the Korean Association of Immunologists T Cell Development and Function Block Symposium II BIOGRAPHY Mizuko Mamura 1992 1992∼1993 1993∼1995 1996∼2000 1999∼2000 2000 2000∼2005 2005∼2007 2007∼2012 2011∼Present 2012∼2014 2014∼Present M.D., Chiba University, School of Medicine, Chiba, Japan Resident, The Second Department of Internal Medicine, Chiba University, School of Medicine, Chiba University Hospital, Chiba, Japan Resident, Yokohama Rosai Hospital, Kanagawa, Japan Ph.D. Student, Teaching Assistant, Laboratory of Allergy and Clinical Immunology, The Second Department of Internal Medicine, Chiba University, School of Medicine, Chiba, Japan Research Fellow, Atopy (Allergy) Research Center, Juntendo University, School of Medicine, Tokyo, Japan Ph.D., Chiba University, School of Medicine, Chiba, Japan Visiting Fellow (200-2003), Research Fellow (2003-2005), Laboratory of Cell Regulation and Carcinogenesis, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA Assistant Professor, Clinical Immunology, Major of Advanced Biomedical Applications, Graduate School of Comprehensive Human Science (Department of Rheumatology and Allergology), University of Tsukuba, Ibaraki, Japan Lee Gil Ya Cancer and Diabetes Institute, Gachon Medical and Science University, Incheon, Korea Adjunct Professor, Department of Molecular Pathology, Tokyo Medical University, Shinjuku, Tokyo, Japan Department of Microbiology, CHA University, Seoul, Korea Internal Medicine, Bundang CHA Hospital, Kyeonggi-do, Korea Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu, South Korea [BS II-3] Dynamic TGF-β Signaling Networks in T Cell Development and Function Mizuko Mamura Department of Molecular Pathology, Tokyo Medical University, Tokyo, Japan, Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu, Korea Transforming growth factor-β(TGF-β) has been characterized as a critical immune regulatory cytokine. TGF-βregulates target gene expression by activation of SMAD transcription factors: SMAD2, SMAD3 (receptor-regulated SMADs) and SMAD4 (common SMAD). Here, we discuss our recent findings on diverse roles of SMADs in the regulation of I> cytotoxic T lymphocytes (CTL) and II> IL-17-producing CD4+ T helper cells (Th17). I> TGF-βsignaling via SMAD4 in combination with SMAD3, but not SMAD2, suppressed the Eomes gene, the essential transcription factor for CTL, and thereby suppressed the differentiation and functions of CTLs. TGF-βantagonism enhanced anti-tumor CTL activity by inducing ubiquitin-mediated degradation of Smad4 in CD8+ T cell-specific manner. II> SMAD2 and SMAD3 oppositely regulated Th17 differentiation, independently of SMAD4. We found that linker-phosphorylated SMAD2 acted as a transcription coactivator of STAT3, whereas C-terminally unphosphorylated SMAD3 acted as a transcription corepressor of STAT3. These findings suggest that SMADs orchestrate T cell development and function in cell-type-specific dynamic signaling networks. The 2014 Fall Conference of the Korean Association of Immunologists 25 Block Symposium II T Cell Development and Function BIOGRAPHY Eun Sook Hwang Eun Sook Hwang is the professor of College of Pharmacy and Graduate School of Pharmaceutical Sciences at Ewha Womans University. Dr. Hwang received her BS degree in Pharmacy from Ewha Womans University and MS and PhD degree in Biochemistry/Molecular Biology from Seoul National University. She worked as a postdoctoral research fellow in Immunology of Harvard School of Public Health. She is a professor at College of Pharmacy and Graduate School of Pharmaceutical Sciences, Ewha Womans University. Dr. Hwang’s laboratory aims to identify the regulatory mechanisms of T helper cell differentiation by T-bet. T-bet undergoes multiple protein modifications and results in suppression of IL-2, induction of IFNγ-producing Th1 cell development, and suppression of Th2 cell differentiation. Dr. Hwang is interested in T-bet’s roles in the regulation of Treg and Th17 cell development and thymic function. Dr. Hwang’s lab is also interested in the modulation of mesenchymal stem cell differentiation by T cell immune system. [BS II-4] Molecular Mechanisms Underlying Thymic Involution Eun Sook Hwang College of Pharmacy and Graduate School of Pharmaceutical Sciences, Ewha Womans University, Seoul, Korea Thymus is an essential endocrine organ for the development and maturation of T cells and the modulation of optimal immune response. Thymus is quite large at the younger age but decreases in size and functional activity upon ageing and inflammation, so-called thymic involution. Thymic involution can result in decreased immune activity and increased susceptibility to microbial infection and tumor progression. Therefore, maintenance of thymus size and function is important for the long-term health. However, the molecular mechanisms underlying thymic involution are largely uncovered. In this study, we aimed to identify the molecules that control thymic involution using starvation and inflammation-induced thymic involution model. We demonstrated that T-bet deficiency resulted in the resistance against starvation-induced thymic involution and overexpression of T-bet decreased thymic cellularity. T-bet increased interferon-γ expression and also enhanced apoptotic cell death in the thymus. More detailed molecular mechanisms remain to be clarified. Keywords: Thymic involution, Starvation, Inflammation, T-bet, Apoptotic cell death 26 The 2014 Fall Conference of the Korean Association of Immunologists Supported by International Vaccine Institute and Seoul National University internal fund. Dendritic Cell and Immune Regulation Block Symposium III BIOGRAPHY Toshiaki Ohteki Dr. Toshi Ohteki is the Professor of Medical Research Institute at Tokyo Medical and Dental University. He received 10th The Japanese Society for Immunology Prize, and was a Principal Investigator of Japanese Society and Technology Agency (JST) Core Research for Evolutional Science and Technology (CREST) Program. His current research focuses on first, differentiation and function of dendritic cells and, second, understanding of tissue homeostasis on the basis of immune cell-tissue stem cell interplay. [BS III-1] Common Dendritic Cell Progenitor (CDP), a Novel Source of Dendritic Cells Toshiaki Ohteki Department of Biodefense Research, Medical Research Institute, Tokyo Medical and Dental University, Japan A major reason people can stay healthy is the protection afforded by the body's immune system. This defense system is essential for performing a social life, and living out one’s life. In the immune system, dendritic cells (DCs), distributed throughout the body as the most powerful antigen-presenting cells, activate immune cells upon viral infection, and maintain immune tolerance to self-antigen under steady-state conditions thereby preventing autoimmune diseases. DCs consist of two major subpopulations, i.e., conventional DCs (cDCs), which have excellent antigen-presenting capacity, and plasmacytoid DCs (pDCs), which have prominent type I interferon (IFN)-productivity, both of which play critical roles in the immune system. Thus, the identification of DC progenitors that give rise strictly to cDCs or pDCs, but not to other hematopoietic cells, could be important in medical applications for treating viral infections and autoimmune diseases. In 2007, in collaboration with Dr. Manz’s group, we identified progenitor cells committed to the DC lineage for the first time. However, these progenitors gave rise to many more cDCs than pDCs, implying that there must be another unidentified type of DC progenitor that serves as a major source of pDCs. After several years’ search, we have recently identified a new type of DC progenitors with prominent pDC developmental potential. In this meeting, I will introduce the new type of DC progenitors and discuss DC differentiation pathways and its possible applications. The 2014 Fall Conference of the Korean Association of Immunologists 27 Hosted by Korean Dendritic Cell Academic Society. Block Symposium III Dendritic Cell and Immune Regulation BIOGRAPHY Florent Ginhoux Florent Ginhoux graduated in Biochemistry from the University Pierre et Marie CURIE, Paris VI and obtained a Masters degree in Advanced Studies in Immunology from the Pasteur Institute, Paris. He then started his PhD in the Immunology Team of GENETHON, Evry and obtained his PhD in 2004 from the University Pierre et Marie CURIE, Paris VI. As a postdoctoral fellow, Florent Ginhoux joined the Laboratory of Miriam Merad in the Mount Sinai School of Medicine (MSSM), New York where he studied the ontogeny and the homeostasis of cutaneous dendritic cell populations, with a strong focus on Langerhans cells. In 2008, he became an Assistant Professor in the Department of Gene and Cell Medicine, MSSM and member of the Immunology Institute of MSSM. He joined the Singapore Immunology Network (SIgN), A*STAR in May 2009 as a Principal Investigator. He is now a Senior Principal Investigator and his laboratory is now focusing on the ontogeny and differentiation of macrophages and dendritic cells (DCs) in both humans and mice. [BS III-2] Dendritic Cell and Macrophage Ontogeny Florent Ginhoux Senior Principal Investigator, Singapore Immunology Network (SIgN), Singapore Dendritic cells (DCs), monocytes and macrophages play crucial and distinct roles in tissue homeostasis and immunity, but also contribute to a broad spectrum of pathologies and are thus attractive therapeutic targets. Potential intervention strategies aiming at manipulation of these cells will require in-depth insights of their origins and the mechanisms that govern their homeostasis. DCs and monocytes arise from common bone marrow (BM) precursor named macrophage-dendritic cell precursors (MDP), branching into exclusively DC- or monocyte-committed progenitors named common dendritic cell progenitors (CDPs) or common monocyte progenitor (cMoPs) respectively. CDPs give rise to plasmacytoid DC and migratory DC precursors termed pre-DCs. Pre-DCs seed tissues where they differentiate into the two major functionally specialized DC lineages, CD8α +/CD103+ DCs and CD11b+ DCs. Recent evidence from our laboratory and others have showed that monocytes do not substantially contribute to all tissue macrophage populations in steady state and inflammatory conditions. Rather certain tissue macrophages in mice are derived from embryonic precursors, are seeded before birth and maintain themselves in adults by self-renewal. In addition, we now provided evidence that commitment to CD8α+/CD103+ DC or CD11b+ DC subsets is imprinted early in the BM. Combining single cell sequencing with conventional transcriptomic analysis, Cytometry by Time-Of-Flight mass spectrometry (CyTOF) and intra-femoral transfer, we identified for the first time DC subset-specific precursors in the BM as well as previously unknown molecular checkpoints for DC lineage commitment as early as the CDP stage. These new insights into the origins of DCs, monocytes and macrophages should aid the rational design of therapies aimed at harnessing the functions of these cells in homeostasis and inflammation and will allow efficient targeting and manipulation during health and disease. 28 The 2014 Fall Conference of the Korean Association of Immunologists Dendritic Cell and Immune Regulation Block Symposium III BIOGRAPHY Myoung Ho Jang My relationship with mucosal immunology started twenty years ago in 1995. At that time, I was involved in a study at the Mogam Biotechnology Research Institute, Korea (one of the WHO Collaborating Centres): screening for the development of new type oral vaccines using nano-particles. However, I had come to recognize my lack of cardinal knowledge required for advancing my project. So, I wrote to Professor Hiroshi Kiyono (Department of Mucosal Immunology, Research Institute for Microbial Diseases, Graduate School of Medicine, Osaka University), a world-renowned researcher in mucosal immunology and decided to take a doctoral course. At Kiyono Laboratory, I devoted myself to research on M (or microfolds) and dendritic cells that ingest antigens in the intestinal lumen. After receiving Ph.D degree, I joined in Professor Miyasaka’s lab. (Immunodynamics, Graduate School of Medicine, Osaka University), my former teacher when I was a post-doctoral researcher. The research on the dynamics of immune cells and chemokines, which had not been clarified at that time, was attractive and interesting, and it developed the basis for the combined study of mucosal immunity and cell kinetics that I have been conducting. I have launched my laboratory in the Osaka University Immunology Frontier Research Center (IFReC), established in October 1st, 2007 as part of the World Premier International (WPI) Research Center Program. It is just like a dream to be able to work as the first Korean principal investigator in immunology field in Japan, and I have been involved in intensive research on dendritic cells and macrophage in the lamina propria of the gut mucosa in IFReC during 5 years. I am now making a concerted effort to generate research findings that may lead to the treatment of cancer and autoimmune diseases and the development of a new oral vaccine in Academy of Immunology and Microbiology (AIM), Institute for Basic Science (IBS), POSTECH. [BS III-3] Intestinal Aldh1a2+CD103+CD8α+ DCs Preferentially Induce Th1 Cells but Not Tregs Bo-Gie Yang, Myoung Ho Jang Academy of Immunology and Microbiology, Institute for Basic Science (IBS), Korea + Intestinal CD103 DCs express retinoic acid (RA)-producing enzyme Aldh1a2 and efficiently promote generation of Foxp3+ Treg cells. These DCs promote expression of gut homing receptors, such as CCR9 and integrin α4β7, on T cells via a RA-dependent mechanism, but are thought to be largely incapable of inducing Th1 cells. However, Th1 cells are abundant in the intestine and IL-12p40 promoter is highly activated in the small intestine under steady-state conditions. Here, we have further divided the CD103+ DC population in the lamina propria (LP) into two different subsets based on CD11b and CD8α expression and found that one of these subsets may be responsible for inducing the Th1 response. Specifically, CD103+CD11b-CD8α+ LP-DCs preferentially produce IL-12 (p35/p40) and IL-12p40 whereas CD103+CD11b+CD8α− LP-DCs secrete IL-23 (p19/p40) upon stimulation with TLR agonists. Consistent with this, CD103+CD11b-CD8α+ LP-DCs preferentially induce Th1 cells whereas CD103+CD11b+CD8α− LP-DCs induce Th17 cells. Interestingly, CD103+CD11b−CD8α+ LP-DCs have lower efficiency to induce Foxp3+ Treg cells compared to CD103+CD11b+CD8α- LP-DCs although both populations imprint gut homing CCR9+ T cells in vitro. We will discuss how Aldh1a2+CD103+CD11b-CD8α+ LP-DCs generate gut-tropic Th1 cells in detail. The 2014 Fall Conference of the Korean Association of Immunologists 29 Block Symposium III Dendritic Cell and Immune Regulation BIOGRAPHY Hyoung-Pyo Kim Dr. Hyoung-Pyo Kim is an Associate Professor of the Department of Environmental Medical Biology at Yonsei University College of Medicine, where he has been since 2008. He obtained his B.Sc., M.S., and Ph.D. degrees from the Department of Microbiology, Seoul National University. In 1998, he moved to Laboratory of Molecular Immunology, National Heart, Lung, and Blood Institute, National Institutes of Health (NIH), as a postdoctoral fellow. Since 2003, he worked as a Research Fellow in the same laboratory. The main focus of his research at NIH was the cytokine systems that critically regulate immune response, and in particular the γc-family cytokines including IL-2 and IL-21. He also delineated a novel link between DNA methylation and FoxP3 expression in regulatory T cells. His current research interests center on transcriptional and epigenetic networks governing dendritic cell lineage specification. He is also dissecting the role of zinc finger protein CTCF in the dynamic change of the nuclear architecture and chromatin function during immune cell differentiation. Additional major interest is in the action of IL-31, a newly discovered cytokine associated with chronic skin inflammation and pruritus. [BS III-4] CTCF Controls Homeostatic Maintenance and Migration of Langerhans Cells in the Skin Tae-Gyun Kim, Hyoung-Pyo Kim Department of Environmental Medical Biology, Institute of Tropical Medicine, Yonsei University College of Medicine, Seoul, Korea Langerhans cells (LCs) are skin-resident dendritic cells (DCs) that orchestrate skin immunity. Regulation of LC homeostasis depends on several extra- and intra-cellular factors. CTCF is a highly-conserved DNA-binding protein that modulates cell differentiation and function through modifying chromatin architectures and subsequent gene expressions. A possible role of CTCF controlling LC homeostasis and function remains to be determined. Herein, by using conditional gene deletion mouse system, we show that CTCF is critically involved in homeostatic maintenance and migration of LCs in vivo. Overall pool of lymphoid and non-lymphoid tissue cutaneous DCs, including LCs was diminished in DC-specific CTCF depletion mice. Another mice with LC-selective CTCF deletion showed a reduced epidermal LC quantity specifically associated with a decreased self-turnover rate. CTCF-depleted LCs exhibited different surface antigen characteristics and distinct activation tendency followed by in vivo hapten challenge. Unexpectedly, CTCF-deficient LCs revealed a functional impairment in migration out of the epidermis. Mice with LC-specific CTCF depletion showed more sustained contact hypersensitivity responses as compared with wild-type littermates. Whole transcriptome analysis of CTCF-deficient LCs revealed differentially expressed gene sets which support cell adhesion and hamper migration. Thus, CTCF promotes LCs to maintain homeostatic pool and efficiently emigrate from the epidermis, revealing an unexpected role of CTCF for this unique cutaneous DC subset. 30 The 2014 Fall Conference of the Korean Association of Immunologists Hot Topics in Transplantation Immunology Block Symposium IV BIOGRAPHY Byungsuk Kwon Byungsuk Kwon is Professor of School of Biological Sciences and Vice-chair of the Foundation of Industry Cooperation at University of Ulsan. He is also joined to University of Ulsan School of Medicine. He received his B.S. degree from Seoul National University and his Ph.D. degree from Wayne State University School of Medicine. Dr. Kwon’s research interest focuses how an immune network formed by CD137-CD13L interactions regulates inflammatory processes in various disease models. Recently, his group has found that CD137L reverse signaling functions as a convergence point for acute tissue inflammation. He is also interested in actions of factors governing tolerance and resistance during infection. [BS IV-1] Integration of the Innate and Adaptive Immunity by CD137-CD137L Bidirectional Signals: Implications in Allograft Rejection Byungsuk Kwon School of Biological Sciences, University of Ulsan, Ulsan, Korea Two-signal models are of use in explaining various types of immune responses. In particular, secondary, so-called costimulatory, signals are critically required for the process of T-cell activation, survival, differentiation, and memory formation. Early studies in rodent models showed that targeting T-cell costimulatory pathways elicits immunological tolerance, paving a basis for the development of costimulatory therapeutics in allograft rejection. However, as the classical definition of T-cell costimulation continues to evolve, simple blockade of costimulatory pathways has limitations in preventing allograft rejection. Furthermore, functions of costimulatory molecules are much more diverse than initially anticipated and beyond T cells. In this lecture, I will discuss CD137-CD137L bidirectional signals as examples showing that two-signals can be applicable to multiple phases of immune responses. The 2014 Fall Conference of the Korean Association of Immunologists 31 Block Symposium IV Hot Topics in Transplantation Immunology BIOGRAPHY Kyung-Mi Lee Kyung-Mi Lee is professor in the Department of Biochemistry and Molecular Biology at the Korea University College of Medicine and project leader of a Korean Global Research Laboratory focused on world-wide collaborative studies in immunobiology field. She obtained B.S. and M.S. degrees from the College of Pharmacy at Seoul National University in 1989 and Ph. D. in the department of Pharmacology and Physiology at the University of Chicago in 1995. She completed postdoctoral training at Harvard Medical School, was appointed as an instructor and research associate professor at the University of Chicago. Since joining the faculty at Korea University in 2003, her research focus in the area of tumor immunology and transplantation has led to the development of novel translational approaches for the treatment of patients with cancer and autoimmune disease. Her laboratory has been involved in cross-disciplinary efforts in biomedical engineering and the development of unique biomaterial platform for immune cell detection, selection, and expansion as a means of enhancing the human immune response both in vitro and in vivo. [BS IV-2] Attenuation of Donor-Reactive T Cells Allows Effective Control of Allograft Rejection Using Regulatory T Cell Therapy Kyung-Mi Lee Department of Biochemistry, Korea University Medical School, Seoul, Korea Regulatory T cells (Tregs) are essential for the establishment and maintenance of immune tolerance, suggesting a potential therapeutic role for Tregs in transplantation. However, Treg administration alone is insufficient in inducing long-term allograft survival in normal hosts, likely due to the high frequency of alloreactive T cells. We hypothesized that a targeted reduction of alloreactive T effector cells would allow a therapeutic window for Treg efficacy. Here we show that preconditioning recipient mice with donor-specific transfusion followed by cyclophosphamide treatment deleted 70-80% donor-reactive T cells, but failed to prolong islet allograft survival. However, infusion of either 5×10(6) Tregs with direct donor reactivity or 25×10(6) polyclonal Tregs led to indefinite survival of BALB/c islets in more than 70% of preconditioned C57BL/6 recipients. Notably, protection of C3H islets in autoimmune nonobese diabetic mice required islet autoantigen-specific Tregs together with polyclonal Tregs. Treg therapy led to significant reduction of CD8(+) T cells and concomitant increase in endogenous Tregs among graft-infiltrating cells early after transplantation. Together, these results demonstrate that reduction of the donor-reactive T cells will be an important component of Treg-based therapies in transplantation. 32 The 2014 Fall Conference of the Korean Association of Immunologists Hot Topics in Transplantation Immunology Block Symposium IV BIOGRAPHY Takaaki Kobayashi Takaaki Kobayashi is Professor of Transplant Immunology, Nagoya University School of Medicine, Nagoya, JAPAN. Kobayashi received his MD in 1985 and PhD in 1993 from Nagoya University School of Medicine in 1985. After he completed the surgical residency for 4 years at Anjo Kosei Hospital and Aichi Cancer Center, he became a clinical fellow in Transplant Surgery at Department of Surgery II, Nagoya University School of Medicine and Department of Transplant Surgery, Nagoya Daini Red Cross Hospital. He made a specialty of transplantation, particularly kidney and liver transplant surgery. Then, he was involved in the research on xenotransplantation, when he worked with Dr. David K.C. Cooper in Oklahoma Transplantation Institute (1994-1995). He assisted the chair person, Dr. Hiroshi Takagi in organizing 5th International Xenotransplantation Association Congress in Nagaya, JAPAN (1999). He has now attempted to produce genetically engineered pigs in collaboration with Prof. Onishi, National Institute of Agrobiological Sciences. His research interests have focused on transplant immunology related to antibody-mediated rejection caused by clinical ABO/HLA incompatible transplantation as well as xenotransplantation. He is a member of The Transplantation Society (TTS: 1997-), International Xenotransplantation Association (IXA: 2001-) and American Society of Transplantation (AST: 2005-). He has served as an elected Councilor of IXA since 2007 and of the Japanese Society for Transplantation since 2003. He was recently appointed a President (2013-2015) of IXA. He has also been the Secretary-General of the Japanese Society for Xenotransplantation since 1997. He has contributed to the development of xenotransplantation as a member of the Editorial Board (2001-) and subsequently as an Associate Editor (2006-) of the official journal of the IXA, ‘Xenotransplantation’. [BS IV-3] Lessons from ABO-incompatible and DSA Positive Renal Transplantation: Potential Strategy to Overcome Chronic Antibody Mediated Rejection Takaaki Kobayashi Department of Transplant Immunology, Nagoya University School of Medicine, Nagoya, Japan Recent advance in immunosuppressive agents has improved the outcome of organ transplantation. We have achieved excellent results with living donor renal transplantation, reaching 5-year graft survival of around 95%. A current problem is that long-term survival has not been extended satisfactorily. It would be essential to take effective measures against chronic antibody mediated rejection (CAMR), which is a main cause of late graft loss and an apparent obstacle to favorable long-term outcome. Our attention has been directed towards blood group ABO-incompatible (ABO-I) and HLA-incompatible (HLA-I) renal transplantation, which can potentially cause antibody mediated rejection (AMR) by anti-A/B and donor specific HLA antibodies (DSA), respectively. Enhanced immunosuppressive regimen including pre-transplant treatment with MMF (for 2 weeks), double filtration plasmapheresis and rituximab or splenectomy (so called desensitization protocol) has provided favorable short and medium-term results. However, one-year protocol biopsy demonstrated that subclinical rejection was observed in 40% of HLA-I, whereas it was only in a few percent of ABO-I, which was comparable to ABO-identical/compatible transplantation without DSA. “Accommodation” which has been termed as a protective condition even in the presence of anti-donor antibody has been proposed as a cause of beneficial results in ABO-I. Our research has focused on identification of “accommodation” mechanisms and development of effective induction ways. Analysis of signaling pathway in endothelial cells revealed that anti-A/B antibody binding could suppress ERK pathway and up-regulate complement regulatory proteins such as CD55 and CD59, which could be related to “accommodation”. Furthermore, we found that inhibition of pro-inflammatory/pro-coagulation response would be necessary, and activation of AMP-activated protein kinase (AMPK) might be one of effective steps to acquire “accommodation”. In the light of clinical findings obtained from renal transplants with preformed or de novo DSA, the risk factors for progression of CAMR such as HLA antibody specificity and complement binding capacity will be discussed. Furthermore, our comprehensive research projects for early diagnosis of CAMR including establishment of in vitro B cell culture system for differentiation into plasma cell and analysis of mRNA/miRNA expression in peripheral blood will be introduced, although they are now in progress. The 2014 Fall Conference of the Korean Association of Immunologists 33 Block Symposium IV Hot Topics in Transplantation Immunology BIOGRAPHY Nayoung Kim Nayoung Kim, Ph.D. is assistant professor at Asan Institute for Life Sciences, Asan Medical Center, Seoul, Korea since 2011. She graduated from Seoul National University, Seoul, Korea where she also received her M.S. degree in Molecular Biology/Immunology. She performed her Ph.D. thesis work on infection immunology at Max-Planck Institute for Infection Biology in Berlin, Germany under Prof. Stefan Kaufmann’s supervision. She worked on NK cell immunology at Dr. Francesco Colucci’s lab, Babraham Institute, Cambridge, UK as a postdoctoral research scientist. Her current research interest is transplant immunology, including NK cell immunology, T cell immunology, liver immunology, ischemia-reperfusion injury, and metabolomics. [BS IV-4] Immune Monitoring of ABO-Incompatible Living Donor Liver Transplant Patients Nayoung Kim Asan Institute for Life Sciences, Asan Medical Center, Seoul, Korea Unlike Western countries, living donor liver transplant surgeries are quite common in Korea and Japan. Recent development of immunosuppressant regimens and monoclonal antibodies has enabled even ABO-incompatible liver transplant (ABOi LT) surgeries. We have recently published a report showing the survival rates and rejection rates were nearly comparable to those of ABO-compatible ones. The only significant problem is biliary stricture in ABOi LT patients. However, there are only a few literatures investigating this problem clinically or lab-based, let alone immunological studies. Therefore we set up this study to monitor immune responses in ABOi LT patients. 34 The 2014 Fall Conference of the Korean Association of Immunologists Autophagy and Regulation of Innate Immunity Block Symposium V BIOGRAPHY Jae Ung Jung Dr. Jae U. Jung is a preeminent expert in the molecular biology of viruses and their gene products as they relate to cell biology, biochemistry and immunology. His research addresses several key biological features of virus-host interactions, with a focus on host immune responses to viruses, mechanisms by which viruses induce tumors, and the ability of viruses to establish life-long infections. He has published over 190 publications on highly peer-reviewed journals and serves as Principal Investigator to several NIH and foundation grants. He is a Fellow of the American Academy of Microbiology and the American Association for the Advancement of Science. In 2012, he received the Ho-Am Prize in Medicine. Prior to his appointment at University of Southern California in 2007, he was a Professor of Microbiology and Molecular Genetics at Harvard Medical School and Chair of Tumor Virology Division at the New England Primate Research Center. He was the first Korean-born scientist to receive tenure at Harvard University. He received his PhD from the University of California, Davis in 1989 and his BS and MS from Seoul National University in Korea in 1982 and 1984, respectively. At USC, Dr. Jung has and will continue to strengthen the basic science efforts and the faculty’s significant contributions in the arena of scientific discovery. He has recruited top-notch investigators, organized multi-investigator program grant award for centers of excellence, and enhanced active core facilities to stimulate the research environment. He continues to integrate programs to bridge gaps between basic science and clinical research, and promote multidisciplinary translation research on the most pressing diseases including AIDS, tuberculosis, flu, autoimmune diseases, and cancers. [BS V-1] Roles of Pattern Recognizing Receptors and Autophagy in Antiviral Immune Response Jae Ung Jung Fletcher Jones Foundation Professor and Hastings Foundation Professor, Chair of Molecular Microbiology and Immunology Department, University of Southern California, Keck School of Medicine, Harlyne J. Norris Cancer Research Tower, 1450 Biggy Street, Los Angeles, CA, USA Understanding the host-viral interaction is an essential step in developing safe and effective anti-microbials against viruses. Since virus is most vulnerable at the early stage of lifecycle, this stage should therefore offer the best opportunity for therapeutic interventions. The early detection of invading viruses by the host depends on a limited number of Pattern Recognizing Receptors (PRRs) that serve as the primary intracellular sensors for viral specific genetic patterns and activate signal transduction cascades, thereby triggering interferon (IFN)-mediated antiviral defense mechanisms. Key virus-detecting PRRs include the nucleic acid-recognizing Toll-like receptors and the cytosolic RNA receptors RIG-I and MDA-5. Specifically, RIG-I has emerged as a key receptor in sensing viruses, including the influenza virus and hepatitis virus C, whereas MDA5 responds to the infection of picornaviruses and noroviruses. On the other hand, the cyclic GMP-AMP (cGAMP) synthetase, called cGAS, is a cytosolic DNA sensor that produces a novel second messenger cGAMP to initiate the STING pathway for IFN production. We demonstrate that the direct interaction between cGAS DNA sensor and Beclin-1 autophagy protein not only suppresses cGAMP synthesis to halt dsDNA stimulation- or herpes-simplex-virus-1 infection-induced IFN production, but also enhances the autophagy-mediated degradation of cytosolic pathogen DNAs to avoid persistent immune stimulation. Specifically, this interaction releases Rubicon, a negative autophagy regulator, from the Beclin-1 complex, activating phosphatidylinositol-3-kinase class III activity and thereby inducing autophagy to remove cytosolic pathogen DNAs. These results indicate that the interaction between cGAS and Beclin-1 shapes innate immune responses by regulating cGAMP production and autophagy pathway, ultimately developing well-balanced immune responses against pathogens. Collectively, this talk will describe a family of host antiviral factors that mediate and elicit host’s innate immunity against viral infections. Keywords: Pattern recognizing receptor, RIG-I, cGAS, Beclin-1, Autophagy The 2014 Fall Conference of the Korean Association of Immunologists 35 Block Symposium V Autophagy and Regulation of Innate Immunity BIOGRAPHY Myung-Shik Lee Dr. Myung-Shik Lee graduated Seoul National University School of Medicine (M.D,), and received Ph.D. from the same University. After post-doctoral training at the Scripps Research Institute, he is Professor in the Dept. of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine. He has been studying cell death and innate immunity in type 1 and type 2 diabetes. He is recently focusing on the topics of autophagy and microbiota in metabolic disorders. He published several papers in Nature Medicine, Immunity, Cell Metabolism, Journal of Clinical Investigation, Journal of Experimental Medicine, PNAS, etc. [BS V-2] Autophagy Insufficiency, Inflammasome and Metabolic Syndrome Myung-Shik Lee Sungkyunwan University, Korea Low-grade tissue inflammation plays an important role in metabolic syndrome and diabetes. Among diverse innate immune receptors, NLRP3, a member of NLR family that, as a constituent of inflammasome complex, can induce maturation of IL-1β, plays a crucial role in tissue inflammation associated with metabolic syndrome and obesity. Autophagy is a process of subcellular membrane rearrangement to form double-membraned autophagosome, and is critical for the proper turnover and function of mitochondria and ER. Autophagy deficiency is a proinflammatory condition characterized by increased activation of inflammasome, which is due to disturbed mitochondrial homeostasis. While the effects of autophagy deficiency on the body metabolism are distinct depending on the location and severity of autophagy deficiency, the role of systemic autophagy insufficiency of physiological relevant range on the body metabolism and tissue inflammatory tone has not been addressed. We generated mice with global haploinsufficiency of an essential autophagy gene (Atg7+/− mice) by breeding Atg7F/F mice with CMV-Cre deleter mice. While Atg7+/− mice did not show metabolic abnormalities, they developed diabetes when bred to ob mice to impose metabolic stress. Atg7+/−-ob/ob mice showed aggravated insulin resistance, increased number of crown-like structure representing adipose tissue inflammation, and enhanced induction of inflammatory genes such as TNFα, IL-6 and pro-IL-1β. Inflammasome activation was more pronounced in adipose tissue of Atg7+/−-ob/ob mice. Intracellular lipid content after lipid loading was increased in autophagy insufficiency, and IL-1βrelease in response to palmitic acid in combination with LPS was increased in macrophages from Atg7+/− mice. Moreover, a decrease in NAD+/NADH ratio, suppressed mitochondrial potential and increase in mitochondrial ROS content after treatment with palmitic acid in combination with lipopolysaccharide was more pronounced in macrophages from Atg7+/− mice. Imatinib or trehalose improved metabolic profile of Atg7+/−-ob/ob mice, which was accompanied by enhanced autophagic flux. These results suggest that systemic autophagy insufficiency could be a factor in the progression from obesity to diabetes by augmenting tissue inflammation through increased inflammasome activation. Autophagy modulators may have therapeutic potential against diabetes associated with obesity and inflammation. 36 The 2014 Fall Conference of the Korean Association of Immunologists Autophagy and Regulation of Innate Immunity Block Symposium V BIOGRAPHY Tatsuya Saitoh Tatsuya Saitoh is the Associate Professor in Department of Host Defense, Research Institute for Microbial Diseases, Osaka University, Japan. He received his B.S. degree in Pharmacology from Chiba University, his M.S. degree in Molecular biology from Hokkaido University, and his Ph.D. degree in Virology from Tokyo Medical and Dental University. After the Ph.D., he moved to Osaka University to work with Dr. Shizuo Akira and became staff scientist in Akira’s lab. Dr. Tatsuya Saitoh focuses on the roles of intracellular degradation systems such as autophagy and proteasome in regulation of innate immune responses. He also studies the roles of organelles such as mitochondria and endoplasmic reticulum in regulation of innate immune responses. Additional interest is to develop an effective treatment for immune-related inflammatory diseases. [BS V-3] Regulation of Innate Immune Response by Autophagy Tatsuya Saitoh Department of Host Defense, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan Innate immunity is induced after sensing microbial components by pattern-recognition receptors and is a first line of host defense against microbes. However, innate immunity is often induced after sensing host-derived stimulatory particles such as monosodium urate (MSU) crystals leading to the development of inflammatory diseases, such as gout. Therefore, better understanding of innate immunity is required for the development of effective therapeutic treatments for infectious and inflammatory diseases. Accumulating evidence has shown that the intracellular clearance system is critically involved in regulation of innate immune response. Notably, the role of autophagy−a degradation system of damaged/old organelles and insoluble protein aggregates−in innate immune response has been highlighted. Here we show recent findings on regulation of inflammatory innate immune response by autophagy. MSU crystal stimulation causes mitochondrial damage and production of reactive oxygen species (ROS), resulting in the activation of inflammatory innate immune response. Autophagy promotes elimination of damaged mitochondria and suppresses ROS-mediated inflammatory response. These findings suggest that autophagy would be an attractive therapeutic target for treatment of immune-related inflammatory diseases. The 2014 Fall Conference of the Korean Association of Immunologists 37 Block Symposium V Autophagy and Regulation of Innate Immunity BIOGRAPHY Eun-Kyeong Jo Prof. Dr. Eun-Kyeong Jo has been leading the medical science research center “Infection Signaling Network Research Center” at Chungnam National University (CNU) since 2007. She gained M.D. degree at 1991 (CNU, School of Medicine), and completed her doctoral thesis in an immunological area of tuberculosis research at CNU. For her postdoctoral training, she changed to the Molecular Immunology Lab at Imperial College London, U.K., and was promoted to professor in Department of Microbiology at CNU School of Medicine since 2008. She has had a long-lasting interest in studying host-pathogen interaction of mycobacterial infection and the innate signaling pathways in mycobacterial infection. She has investigated a novel function of nuclear receptors and autophagy in infection and inflammation. She has won prestigious awards and published many beautiful papers in medical and science fields. [BS V-4] Small Heterodimer Partner and Innate Immune Regulation Eun-Kyeong Jo Department of Microbiology and Infection Signaling Network Research Center, Chungnam National University School of Medicine, Daejeon, Korea When an inflammatory state is excessive or unregulated, local and systemic damage to host tissues can result in pathologic or deleterious status. Currently, accumulating evidence has revealed that several members of nuclear receptor superfamily regulate immune and inflammatory responses through specific modes of interaction, and/or regulation of gene expression to exert for maintaining homeostasis in the body. Our previous studies showed that the orphan nuclear receptor SHP (small heterodimer partner) is an intrinsic negative regulator of TLR-triggered inflammatory responses. SHP had dual regulatory functions in a canonical transcription factor NF-κB signaling pathway, acting as both a repressor of transactivation of the NF-κB subunit p65 and an inhibitor of polyubiquitination of the adaptor TRAF6. Fenofibrate, a drug for reducing cholesterol and triglyceride levels, significantly blocked endotoxin-triggered inflammatory signaling responses via SHP. Our current study for the SHP in regulation of NLRP3 inflammasome activation will be also discussed. SHP deficiency resulted in an increased secretion of interleukin (IL)-1βand IL-18, and excessive pathologic responses typically observed in vitro and in vivo. Collectively, these data suggest that the SHP-inducing drugs pave the way for novel therapies for diverse inflammatory settings through targeting SHP. Keywords: Orphan nuclear receptor, Small heterodimer partner (SHP), Inflammation, Innate immunity, Inflammasome 38 The 2014 Fall Conference of the Korean Association of Immunologists Mucosal Immunology and Vaccine Block Symposium VI BIOGRAPHY Dipayan Rudra I did my Bachelors and Masters from the University Of Calcutta, India, in Chemistry and Biochemistry respectively, following which I completed Ph.D. in biological sciences from the Department of Cell Biology in Albert Einstein College of Medicine, Bronx, New York under the supervision of Dr. Jonathan Warner. During my post doctorate years I worked with Dr. Alexander Rudensky in Memorial Sloan Kettering Cancer Center, New York. Since September 2013 I have joined as an Assistant Investigator in the Academy of Immunology and Microbiology (AIM) at the Institute for Basic Science (IBS), POSTECH campus. Research in our laboratory is focused on the biology of Regulatory T cells. [BS VI-1] Immuno-Modulatory Transcriptional Complexes in Regulatory T Cells Dipayan Rudra Academy of Immunology and Microbiology/POSTECH Campus, Institute for Basic Science (IBS), Korea Regulatory T (Treg) cells comprise an indispensable population of our immune system, and due to their well-established immune-regulatory functions, hold enormous therapeutic promise for autoimmunity and cancer immunotherapy. Moreover, the existence of a lineage specific transcription factor in Treg cells, Foxp3, makes them an attractive model for studying basic aspects of cellular differentiation and function. My research is focused on dissecting the molecular determinants accounting for Foxp3-mediated gene expression, with a long-term goal of translating this knowledge into potential therapeutic interventions. Recently, using a mass-spectrometry based proteomic approach; we generated an exhaustive list of Foxp3-associated proteins involved in establishing Treg cell-specific gene regulatory networks. The biochemical and mass-spectrometric analyses revealed that Foxp3 forms multiprotein complexes of 400-800 kDa or larger and identified 361 associated proteins, ∼30% of which are involved in the regulation of transcription. Notably, Foxp3 binds and directly regulates expression of a large proportion of the genes that serve as its co-factors. In reciprocation, some of the sequence-specific transcription factors that serve as Foxp3 binding partners facilitate Foxp3 expression. Functional analysis of Foxp3 cooperation with at least two of its associated partners provides further evidence for a cooperative network of transcriptional regulation afforded by Foxp3 and its associates to control distinct aspects of Treg cell biology. The relevance of these findings in light of the current research focus of our laboratory will be discussed. The 2014 Fall Conference of the Korean Association of Immunologists 39 Block Symposium VI Mucosal Immunology and Vaccine BIOGRAPHY Joon Haeng Rhee A graduate of Chonnam National University Medical School and received PhD from the same university. I have been working on molecular microbial pathogenesis and vaccine biology. For the molecular microbial pathogenesis studies, our laboratory has been observing the V. vulnificus-host interactions using various molecular and cellular microbiological tools. We reported the whole genome sequence of CMCP6, which became the most widely used standard strain in the V. vulnificus research field. We identified a RTX toxin as the culprit of deadly host-killing mechanism in the V. vulnficus infections. Vaccine study was first started aiming the high mortality V. vulnificus infections. During the vaccine research, our team came across the finding that a flagellin protein of V. vulnficus has an excellent mucosal adjuvant effect in late 1990s, which was later proved by our group and others to be mediated by the TLR5 signaling. Currently our laboratory is studying the basic science and applications related to the flagellin-TLR5-mediated immune modulation. [BS VI-2] A New Therapeutic Approach to Asthma Utilizing the TLR5 Signaling Pathways Joon Haeng Rhee Department of Microbiology and Clinical Vaccine R&D Center, Chonnam National University Medical School, Gwangju, Korea Toll-like receptor (TLR) ligands are considered attractive adjuvants for vaccines and immunotherapy. TLR stimulation leads to activation of innate and subsequently modulates adaptive immune responses. Flagellin is the cognate ligand for TLR5 of host cells. Previously, we have shown that flagellin has a unique immunomodulating activity especially in the mucosal immune compartment. Allergic asthma is an immunologic disease characterized by eosinophilic airway inflammation. While the ‘hygiene hypothesis’ suggests microbial infections could subvert allergies, microbial products were reported to promote allergic asthma. Low-level flagellin, the TLR5 ligand, was reported to prime allergic responses to indoor allergens. However, we found that therapeutic doses of flagellin (FlaB) together with allergens suppress experimental. FlaB/ovalbumin (OVA) mixture ameliorated experimental asthma by inhibiting pathogenic Th1/Th2/Th17 responses while generating regulatory DCs (DCreg) and Treg cells. Adoptive transfer of FlaB/OVA mixture-induced DCs inhibited asthma while depletion of CD25+ cells eliminated the inhibiting effect of the mixture. Similar inhibitory effect of FlaB was observed in combination with house dust mite (HDM) in HDM-induced experimental asthma. In peripheral blood from HDM-sensitive asthma patients, FlaB treatment induced DCreg, which subsequently induced HDM-specific Foxp3+ Treg cells in a lymphocyte co-culture while inhibiting Th1/Th2 responses in an IL-10-dependent manner. These findings collectively suggest that flagellin could be employed as a tolerogenic adjuvant in allergen-specific immunotherapy (SIT) of allergic asthma. 40 The 2014 Fall Conference of the Korean Association of Immunologists Mucosal Immunology and Vaccine Block Symposium VI BIOGRAPHY Mitchell Kronenberg Mitchell Kronenberg received an undergraduate degree from Columbia University and a Ph.D. from the California Institute of Technology in 1983, where he continued as a postdoctoral fellow in the laboratory of Dr Leroy Hood. Dr. Kronenberg served on the faculty of the UCLA School of Medicine in the Department of Microbiology and Immunology from 1986-1997. He moved to the La Jolla Institute for Allergy & Immunology in 1997, and was appointed President in 2003. He is responsible for the leadership and overall administration of the Institute, which has grown to 23 faculty and more than 375 employees since its establishment in 1988. He also is an Adjunct Professor of Biology at UC San Diego. Dr. Kronenberg has co-authored more than 310 total publications, and his awards include an NIH Merit Award; Kroc Distinguished Professor in Medicine and Immunology at UC Davis, a Burroughs Wellcome Fund Visiting Professor at Harvard University., and numerous named or distinguished lectureships He is an Institute for Scientific Information “Highly Cited Scientist” and he has also chaired several international meetings, such as the Gordon Conference of Immunobiology and Immunochemistry and a Keystone Symposium, and he participated as a member of the Japan-U.S. Cooperative Medical Board for Immunology. Dr. Kronenberg recently served as Deputy Editor of the Journal of Immunology, and serves on the Council of the American Association of Immunologists. Dr. Kronenberg’s research interests include antigen recognition by T lymphocytes, mucosal immunology, and inflammatory and autoimmune disease models. Some of his recent efforts have concentrated on Natural Killer T (NKT) cells, a unique T lymphocyte subpopulation that combines features of innate and adaptive immunity in responding to bacterial glycolipids. Another area of research interest concerns the role of TNF family ligands and receptors in regulating mucosal immune responses. Mitchell has collaborated for many years with Dr. Hilde Cheroutre, who is also a member of the La Jolla Institute for Allergy & Immunology faculty. Their joint work covers several areas, including the specificity and function of intraepithelial lymphocytes, CD8 T cell memory and the regulation of mucosal inflammation. [BS VI-3] HVEM: a TNF Family Receptor That Controls Diverse Aspects of Mucosal Immunity Mitchell Kronenberg La Jolla Institute for Allergy & Immunology, USA The herpes virus entry mediator (HVEM) is a TNF super family receptor that binds multiple ligands, including TNF member LIGHT and Ig super family members BTLA and CD160. We have explored the role of HVEM in epithelial cells. During Citrobacter rodentium infection by oral gavage, Hvem−/− mice had impaired colonic epithelial responses, resulting in higher bacterial burdens, inflammation and increased mortality. HVEM stimulation induced epithelial responses by a novel signaling pathway, with NIK-dependent Stat3 activation acting downstream of HVEM, resulting in the expression of genes important for mucosal immunity. HVEM is engaged by CD160, which is highly expressed by intraepithelial lymphocytes of the intestine. HVEM is also important for innate epithelial responses in the lung, but in this case the ligand for HVEM is BTLA. A type of innate lymphoid cells (ILC) called ILC3, also expresses HVEM. ILC3 are characterized by the production of IL-22, and HVEM expression by these cells is required for their optimal production of this cytokine. In the absence of HVEM expression by ILC3 in conditional HVEM knockout mice, constitutive IL-22 production is reduced with corresponding changes in the microbiota and a resulting increase in mucosal Th17 cells. Therefore, HVEM expression by different cell types in the intestine has important effects on both innate and adaptive immunity. The 2014 Fall Conference of the Korean Association of Immunologists 41 Block Symposium VI Mucosal Immunology and Vaccine BIOGRAPHY Chen Dong Dr. Dong did his Ph.D. training with Dr. Max Cooper and his postdoctoral study was conducted in the lab of Dr. Richard Flavell. He served as the Director of the Center for inflammation and Cancer at the University of Texas MD Anderson Cancer Center and is currently the Director of the Institute for Immunology at Tsinghua University. Dr. Dong’s research is to understand the molecular mechanisms whereby immune and inflammatory responses are normally regulated, and to apply this knowledge to the understanding and treatment of autoimmunity and allergy disorders as well as cancer. The work from Dr. Dong’s group has led to the discoveries of Th17 and T follicular helper (Tfh) cell subsets in the immune system and elucidation of their biological and pathological functions. Dr. Dong has over 170 publications and the honors he has received include the 2009 American Association of Immunologists BD Bioscience Investigator Award and election of fellow, the American Association for the advancement of Science in 2011. In 2014 Professor Dong is awarded as Highly Cited Researcher by Thomson Reuters based on publications in 2002-2012. [BS VI-4] IL-17 Family Cytokines in Mucosal Inflammation and Cancer Chen Dong Institute for Immunology, Tsinghua University, Beijing, China IL-17 cytokine family consists of six members, IL-17 (also known as IL-17A), IL-17B, IL-17C, IL-17D, IL-17E (also called IL-25) and IL-17F. These cytokines have been shown to promote inflammatory responses. IL-17 and IL-17F are produced by a novel subset of T cells, called TH17 cells. IL-17 has been well studied and shown to drive inflammation and regulate cancer development in vivo. IL-25 is associated with allergic responses and our previous study has indicated an important function of this cytokine in regulation of asthma disease. Recent work has suggested even broader function of IL-25 in inflammatory diseases. Further studies on IL-17 cytokines may help understand the inflammatory diseases and suggest novel targets for these diseases. 42 The 2014 Fall Conference of the Korean Association of Immunologists Clinical Immunology Block Symposium VII BIOGRAPHY Richard Bucala Dr. Bucala obtained a BS and MS degree from Yale University, and an MD/PhD from the combined program at Rockefeller University. After a fellowship at the Pasteur Institute in Paris, Dr. Bucala completed his residency training at the Brigham and Women’s Hospital and Harvard Medical School, and then returned to Rockefeller to pursue post-doctoral studies together with clinical fellowship training in Rheumatology at The Hospital for Special Surgery. Dr. Bucala joined the faculty at Rockefeller University and then served as Scientific Director of the Picower Institute for Medical Research. He was recruited to the Yale Faculty in 2002. The laboratory’s main research focus is on MIF-family cytokines and their receptors. The Bucala group discovered MIF to regulate glucocorticoid immunosuppression, which opened novel approaches to immunotherapy, and it went on to clone the two-component MIF receptor and to identify function polymorphisms in the MIF gene, which show significant population stratification. Depending on the nature of the immune, infectious, or invasive provocation, MIF may be either protective of disease or contribute to severe clinical manifestations. The laboratory has developed inexpensive biochips to determine MIF polymorphisms in field settings; these have had application for global infectious disease studies in malaria and tuberculosis. The laboratory also is centrally involved in developing MIF-based therapies tailored to an individual’s genetic makeup. A humanized anti-MIF is in phase I clinical testing for SLE and for solid tumors, and an anti-MIF receptor antibody is in phase II studies for leukemias/lymphomas. Collaborative work in structure-based drug design has produced the first small molecule cytokine antagonists using the MIF/MIF receptor as a targeted interaction. One such orally bioavailable MIF antagonist is in advanced pre-clinical testing for autoimmunity. Small molecule MIF agonists also have been discovered in this program and are being studied for application as immune adjuvants and in ischemic cardioprotection. The research program also has expanded to encompass the recently characterized MIF homolog, DDT, as well as the orthologous MIF-like proteins produced by the parasitic pathogens responsible for malaria, leishmaniasis, and hookworm infection. Collaborative studies in Africa focus on MIF's role in the development of global infectious diseases. The Bucala group conducts clinical studies at the Macha Mission Hospital in Zambia to examine the frequency of different MIF genetic polymorphisms in an effort to understand why severe malaria develops in certain children. Additional work is underway in tuberculosis and in Leishmaniasis. A second major interest is the fibrocyte, which is a circulating connective tissue cell first isolated by the Bucala group and that contributes to the pathogenesis of different systemic and organ-specific fibrosing disorders. Fibrocytes are prognostic biomarkers for interstitial lung disease and a fibrocyte directed therapy (rhPTX-2) is currently in phase II clinical evaluation. At Yale, Dr. Bucala also teaches clinical medicine and lectures in the Schools of Medicine, Public Health, and in Yale College on the topics of inflammation, autoimmunity, and malaria. [BS VII-1] Approaching the Gene-specific Therapy of Autoimmunity Richard Bucala Yale School of Medicine, USA Our research focuses on the mechanisms by which host immunity converts from a protective response to one producing autoimmunity. Beginning with the cloning of the cytokine MIF and the definition of its unique neuroendocrine role in the host response, our laboratory has pursued structure-function, signal transduction, and genetic studies. MIF’s role in autoimmunity has been underscored by our discovery of functional polymorphisms in the MIF promoter, and human genetic studies continue to define the contribution of commonly occurring MIF alleles to the susceptibility or clinical severity of many inflammatory and infectious diseases. Currently, we are leading efforts to develop MIF-based therapies tailored to an individual’s genetic makeup. A neutralizing anti-MIF antibody has been humanized for clinical evaluation in SLE and in cancer, and collaborative work in structure-based drug design has led to orally active, small molecule MIF antagonists that are in advanced pre-clinical testing. MIF agonists also are under development for application as immune adjuvants and in ischemic tissue protection. Finally, the function of the MIF-like genes expressed by the parasitic pathogens responsible for malaria, leishmaniasis, and hookworm disease are under investigation for their ability to suppress immunologic memory. We anticipate that these efforts will provide a means to selectively intervene in a key immunologic pathway, with the ultimate objective of re-setting an individual’s genetic susceptibility to disease. The 2014 Fall Conference of the Korean Association of Immunologists 43 Block Symposium VII Clinical Immunology BIOGRAPHY Young Mo Kang Young Mo Kang is the professor of Internal Medicine (Rheumatology) at the Kyungpook National University (KNU) School of Medicine and Kyungpook National University Hospital. He has served as an Associate Dean of Research Programs and as the first chairman of Institutional Animal Care and Use Committee (IACUC) of KNU with establishment of Regulations for KNU IACUC. He is currently the Chief Professor of Department of Internal Medicine and Director of Cell & Matrix Research Institute at KNU. He is passionate about networking clinical researchers and basic scientists who have interest in trans-disciplinary cooperation for arthritis research. Nationwide, He got actively involved in the establishment of Korean Society of Synovitis Research and served as the first president. As a clinical researcher, he has focused on translating basic research to clinical diseases and vice versa. He has been studied the cell-matrix interaction within inflammatory microenvironment where circulating inflammatory cells are recruited through activated endothelium and guided by matrix such as βig-h3. Based on the understanding of critical role of extracellular matrix for cell dynamics that drives pro-inflammatory niche, his lab has focused on the development of new therapeutic agents including matrix metalloproteinase1-cleavable peptide based on βig-h3 fragments. [BS VII-2] βig-h3 and Its Therapeutic Implications in Rheumatoid Arthritis Young Mo Kang Department of Internal Medicine (Rheumatology), Kyungpook National University School of Medicine, Daegu, Korea Extracellular matrix (ECM) provides the mechanical scaffolding and also takes part in the adhesive interaction of cells via integrins. βig- h3 (TGF-beta inducible gene-3), abundantly expressed in the synovial tissues and fluid of patients with rheumatoid arthritis (RA), has regulatory roles in cellular growth, differentiation, adhesion, migration, and angiogenesis. βig-h3 play an importatnt role in the recruitment of memory T cells, which are expressing high level of α5β1 integrin within the inflamed synovial tissues. Extravasating T cells within synovial tissues first come in contact with ECM proteins including βig-h3 and then are activated via cell-to-cell contact and/or by inflammatory mediators in the context of ECM. The interaction mediated by βig-h3 involves multiple cell adhesion motifs within the fas-1 domains that can bind to different cell types through distinct integrins. The selection of the integrin may depend on the activation state of the integrins in each cell type. Furthermore, different ECM proteins may be engaged in the interaction with cells using a common integrin, implicating the possible application of βig-h3 derivatives for blocking cellular adhesion on other ECM proteins. Based on the findings that βig-h3 is abundantly expressed in rheumatoid synovium, and matrix metalloproteinases (MMP) play important roles in the pathogenesis of RA, a proof-of-concept design of MMP-cleavable composite peptide was developed and showed a markedly improved therapeutic efficacy in chronic inflammatory arthritis, providing a new expandable strategy for enhancement of the efficacy of two different active molecules in RA. 44 The 2014 Fall Conference of the Korean Association of Immunologists Clinical Immunology Block Symposium VII BIOGRAPHY Jae Hee Cheon Prof. Cheon graduated from Seoul National University College of Medicine in 1996. He obtained Ph.D. at Seoul National University College of Medicine in 2006. He is now working at the department of Gastroenterology, Internal Medicine, Yonsei University College of Medicine. His major clinical and research activities have focused on inflammatory bowel disease and its related mucosal immunology and genetics. He has published more than 100 original and review papers in international SCI(E)Journals, especially regarding IBD, intestinal Behcet’s disease, and clinical endoscopy. Currently, he is the project leader of 3 national scientific research projects for genetics and immunology of IBD. [BS VII-3] Genetic and Immunologic Aspects of Inflammatory Bowel Diseases Jae Hee Cheon1,2 1 Department of Internal Medicine and Institute of Gastroenterology, Yonsei University College of Medicine, Seoul, Korea, 2 Avison Biomedical Research Center, Yonsei University College of Medicine, Seoul, Korea Inflammatory bowel disease (IBD) is a chronic intestinal inflammatory condition stemming from unidentified causes, including Crohn’s disease and ulcerative colitis. Although its precise pathogenesis is still unknown, IBD is believed to occur as the result of interactions among genetic, bacterial, and immunological factors that induce intestinal inflammation. Genetic studies are gaining popularity to answer this question since DNA underlies nearly every aspect of human health, including function and dysfunction. Individual differences in IBD susceptibility may be best explained by a detailed picture of how genes work together and interact with environmental factors. Therefore, genetic studies have opened a window into the complex biology of IBD. Various candidate genes for IBD have been discovered through genome-wide association studies (GWAS) or candidate gene approaches. The research efforts of IBD genetics, especially by means of GWAS, have led to the identification of at least 160 genetic loci. Yet, functional studies for a direct pathogenetic role in IBD have been well established for only three genetic polymorphisms related to NOD2, IL23/Th17, and autophagy. Moreover, there is increasing evidence of global variation in IBD susceptibility genes, especially regarding significant differences in specific variants of the same gene, and polymorphism frequency between racial and ethnic groups. Dynamic interactions between commensal microflora and the gut immune system are critical for establishing intestinal homeostasis. In the literature, researchers have primarily focused on understanding dysregulated mucosal immune homeostasis as a main pathogenetic mechanism of IBD. These studies generally regard the dynamic imbalance between intestinal microbes and the host mucosal defensive system as playing an essential role in the initiation and pathogenesis of IBD. Over the last few decades, the majority of studies on commensal microbiota have focused on gut bacteria. However, gut microorganisms consist of not only bacteria but also fungi or viruses. In a recent paper, fungi in the gut interacted strongly with the intestinal immune system through the innate immune receptor Dectin-1, which was shown to regulate colitis severity. Moreover, an interaction between a specific virus infection and a mutation in Atg16L1 was shown to induce intestinal pathologies in mice, providing a specific example of how a virus-plus-susceptibility gene interaction can determine the phenotype of hosts carrying common risk alleles for IBD. Here, we demonstrate that intestinal viruses can induce intestinal homeostasis via Toll-like receptors 3 and 7. Overall, fungi and viruses, as well as bacteria, in the gut interact strongly with the gut immune system and might essentially change how we think about the relationship between gut microflora and IBD. The 2014 Fall Conference of the Korean Association of Immunologists 45 Block Symposium VII Clinical Immunology BIOGRAPHY Yoon-Keun Kim Yoon-Keun Kim (MD, PhD) graduated in Medicine from Seoul National University in 1987. After training in residency at the Seoul National University Hospital and military service, he had researched and published several excellent articles about spider mite including citrus red mite, which is very important allergen in Korea causing allergic rhinitis and asthma, for the first time in the world. He had got the extinguishing award with this research work from Seoul National University Alumni in 1999. And since then he did his research in the Department of Allergy and Clinical Immunology, Seoul National University College of Medicine as an assistant professor. He came to the Division of Pulmonary and Critical Care at Yale University as a visiting assistant professor in 2002. With Dr Jack A. Elias who is outstanding worldwide researcher in the field of allergy and respiratory medicine, he had done several research projects on the pathogenesis of asthma using mouse models for two years, and they published the epoch-making articles in the journal ‘Science’ and ‘Nature Medicine’. Coming back to Korea, he has actively working about the interesting and very important mechanisms in the pathogenesis of asthma, ‘new paradigm in the immunopathogenesis of asthma’. He demonstrated that non-eosinophilic asthma is induced by both Th1 and Th17 immune responses, whereas eosinophilic asthma by Th2 immune response. Moving his position to the Department of Life Science, POSTECH, he has performed the research with enthusiasm and extended his interests to the pathophysiology of immune-based inflammatory diseases and translational research covering cutaneous, gastrointestinal, and cardiovascular diseases in addition to the respiratory diseases. Especially, his recent research outcomes regarding extracellular vesicles are highly creative and practical in the context of developments of biomarkers, preventive devices, and therapeutics for the management of diverse diseases. These have its novelty in the fields, and this is reflected on his recent patent submissions. His two recent publications that extracellular vesicles from Staphylococcus aureus induce atopic dermatitis and that extracellular vesicles from intestinal commensal bacteria induce systemic inflammation, provide new paradigm of pathogenesis of immune-based inflammatory diseases with unknown etiology. Regardless of his publications, his research outcomes have the very high potential to be applied in the management of immune-based inflammatory diseases and cancers. These coming outcomes will benefit enormously to the progress of the science and contribute to the human health, and even to the economy in this country. [BS VII-4] Role of Bacteria-derived Extracellular Vesicles on the Pathogenesis of Chronic Obstructive Airway Disease Yoon-Keun Kim Director, Ewha Institute of Convergence Medicine and Professor, Ewha Womans University Medical Center, Seoul, Korea The role of infectious agents in the etiology of inflammatory diseases once believed to be non-infectious is increasingly being recognized. Many bacterial components in indoor dust can evoke inflammatory pulmonary diseases. Especially, we recently found that bacteria-derived EVs in indoor dust are pathophysiologically related to chronic inflammatory airway diseases which are characterized by neutronphilic inflammation. To elucidate pathogenic bacteria or bacteria-derived EVs, we performed metagenomic analysis of bacteria and bacteria-derived EVs in indoor dusts. Microbiota compositions in indoor dust revealed the presence of both gram-negative and gram-positive bacteria, with most sequences (＞90%) related to just five genus: Pseudomonas (61.6%), Enterobacter (13.6%), Acinetobacter (7.0%), Leclercia (4.5%), and Staphylococcus (0.9%). E. coli is a model organism of gram-negative Enterobacteriaceae. We evaluated the role of E. coli-derived EVs on the development of lung pathology. To induce lung pathology, E. coli EVs were administrated intranasally into the airways of 6 weeks-old mice six times on days 0, 1, 7, 8, 14 and 15. Pulmonary inflammation, emphysema, and immunologic parameters were evaluated 48 h after the final EVs administration. E. coli-derived EVs were present in indoor dust. Repeated inhalation of E. coli-derived EVs caused neutrophilic inflammation and emphysema in a dose-dependent manner. These phenotypes were accompanied by the production of both Th1 and Th17 cells. Additionally, emphysema induced by E. coli-derived EVs was partially eliminated by the absence of IFN-gamma or IL-17. Taken together, E. coli-derived EVs induce lung emphysema via both IFN-gamma and IL-17 dependent pathways, and EVs in indoor dust, especially derived from Gram-negative bacteria, appear to be an important causative agent in the pathogenesis of neutrophilic asthma and/or emphysema. 46 The 2014 Fall Conference of the Korean Association of Immunologists Hosted by BK21+ Frontier Pharmacy Leaders, Chungbuk National University. KAI-AKIA Joint Symposium Block Symposium VIII BIOGRAPHY Chang Hwan Kim He received his B.S. and M.S. degrees from KAIST before he joined LG biotech as a scientist. Then, he received his PhD degree at Indiana University School of Medicine and moved to Stanford University for research fellowship. In 2002, he became a professor of Immunology at Purdue University and is currently the section head of Microbiology and Immunology in the Department of Comparative Pathobiology and the Weldon School of Biomedical Engineering at Purdue University. He has been studying immune cell trafficking and differentiation with the major focus on CD4 T cells. He discovered the migration of hematopoietic stem and progenitor cell migration to the chemokine SDF-1. He found a group of T cell subsets migrating to B cell follicles, now referred to as T-FH cells or GC-T cells, which are specialized in helping B cells. His group found the function of retinoic acid in inducing gut Tregs for immune tolerance. Recently, his group has elucidated the important roles of a group of gut microbial metabolites called short-chain fatty acids in regulating immunity and immune tolerance through epithelial cells and T cells. [BS VIII-1] Regulation of Mucosal Immunity by the Gut Microbial Metabolites Short Chain Fatty Acids Chang Hwan Kim Comparative Pathobiology and Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, USA Microbial metabolites such as short chain fatty acids (SCFAs) are highly produced by gut commensal bacteria in the intestine and profoundly regulate the immune system. Acetate, propionate and butyrate are the major SCFA species and produced as microbial fermentation products from soluble carbohydrate fibers in diets. The intestinal immune system is in a unique tissue environment where one layer of epithelial cells separates the tissue environment from foreign antigens and microbes. The gut tissue is highly enriched with functionally specialized innate and adaptive immune cells including regulatory and effector T cell subsets. We studied the function of SCFAs and their receptors in regulating the two functionally important cell types in the intestine: epithelial cells and T cells. For the first part, I will discuss the functions of SCFA receptors (GPR41 and GPR43) in regulating epithelial inflammatory responses to commensal bacteria following gut barrier breakage or infection with enteric pathogens. We found that SCFAs condition intestinal epithelial cells to more efficiently produce chemokines and cytokines for recruitment of leukocytes and effector T cells to the intestine. This is mediated through activation of ERK1/2 and p38 mitogen-activated protein kinase (MAPK) signaling pathways following GPR41 or GPR43 triggering. The second part is about the function of SCFAs in regulating T cell activity. We found that SCFAs promote the induction of T cells producing IL-17 (Th17), IFN-γ (Th1), and/or IL-10 (Tr1) depending on cytokine milieu. Selective induction of these T cell subsets in appropriate situations is important for regulating both immunity and immune tolerance. The SCFA effect on T cells is mediated through inhibition of histone deacetylase (HDAC) and regulation of the mTOR pathway. Taken together, the results indicate that SCFAs profoundly regulate the gut immune system in both positive and negative ways and this is important for promotion of immunity to pathogens but suppress unwanted inflammatory responses. The 2014 Fall Conference of the Korean Association of Immunologists 47 Block Symposium VIII KAI-AKIA Joint Symposium BIOGRAPHY You-Me Kim Dr. You-Me Kim is an assistant professor in the Division of Integrative Biosciences and Biotechnology (IBB) of Pohang University of Science and Technology (POSTECH) in Korea. She received her B.S. and M.S. degree from Seoul National University and her Ph.D. degree in Molecular Pharmacology and Structural Biology from Thomas Jefferson University in U.S.A. After 5 years of her postdoctoral training at Harvard Medical School and Whitehead Institute, she worked at Novartis Institutes for Biomedical Research until joining POSTECH in 2009. Dr. Kim’s research focuses on elucidating the cellular and molecular mechanisms of the immune responses. Her main research topics include signaling and regulation of immune receptors such as Toll-like receptors and B cell receptors. Recently, she also started to study pathogenesis of systemic lupus erythematosus and regulation of type I interferon pathways. The ultimate goal of her research is to provide the framework for the therapeutic intervention of inflammatory and autoimmune diseases. [BS VIII-2] Intracellular Trafficking of Toll-Like Receptors You-Me Kim Division of Integrative Biosciences and Biotechnology & Department of Life Sciences, Pohang University of Science and Technology POSTECH Biotech Center, Pohang, Korea Toll-like receptors (TLRs) are one of the most important innate immune receptors. They directly recognize pathogen-associated molecular patterns (PAMPs) and induce immediate defense responses against invading microbes for the host. More recently, endogenous molecules derived from stressed or dying host cells, such as DNA, RNA, chaperones and nuclear proteins, have also been shown to activate TLRs. Over-activation of TLRs by these danger-associated molecular patterns (DAMPs) often contribute to various immune disorders. TLRs are divided into two groups based on their cellular localization patterns; TLR1, 2, 4, 5, 6 are localized on the cell surface whereas the nucleotide-sensing TLRs such as TLR3, 7, 8, 9 are found intracellularly in the majority of cell types. We have previously demonstrated that the polytopic membrane protein UNC93B1 physically interacts with the nucleic acid-sensing TLRs and deliver them from the ER to endolysosomes wherein the receptors bind internalized nucleic acids and initiate signaling. In the UNC93B1 mutant cells, the nucleic acid-sensing TLRs are unable to exit the ER and consequently fail to signal. In addition, we found that acidic amino acid residues in the luminal juxta-membrane region of the nucleic acid-sensing TLRs are essential for binding to UNC93B1 and for the proper localization in endolysosomes. Based on the presence of the juxta-membrane acidic residues, we also discovered that TLR5, expressed on the cell surface and therefore regarded to traffic in an UNC93B1-independent manner, also requires UNC93B1 for localization and signaling at the plasma membrane. 48 The 2014 Fall Conference of the Korean Association of Immunologists Hosted by BK21+ Frontier Pharmacy Leaders, Chungbuk National University. KAI-AKIA Joint Symposium Block Symposium VIII BIOGRAPHY Minsoo Kim Unlike cells within solid tissues, circulating leukocytes relocate during the course of immune reactions and dynamically adhere and de-adhere to cells of the vasculature and to other immune cells, as well as to components of the extracellular matrix. A subset of integrins are expressed on leukocytes and play a major role in regulating leukocyte adhesion and recruitment to damaged or infected tissues during inflammatory responses. Under a wide range of pathologic conditions, integrin activation is mis-regulated resulting in abnormal leukocyte trafficking, and direct damage to the vasculature and the underlying tissue making leukocyte integrins a promising therapeutic target for anti-inflammation therapy. Our current research focuses on integrin & chemokine-mediated leukocyte migration during inflammation. By employing advanced imaging techniques including two-photon intravital microscopy, FRET, 3D confocal-image reconstruction, and TIRFM (total internal reflection fluorescence microscopy), we are trying to improve our understanding of the supramolecular/architectural properties of leukocyte integrins with particular emphasis on their relationship with leukocyte migration. In addition, we are interested in visualizing dynamic interactions of intracellular signaling molecules and identifying new molecules that regulate integrin activation during the leukocyte chemotaxis. [BS VIII-3] Neutrophils Provide Local Chemokine Cues to Regulate CD8 T Cell Responses to Influenza Minsoo Kim Center for Vaccine Biology & Immunology, Department of Microbiology & Immunology, Rochester, NY, USA Upon virus infection, precise trafficking of activated effector T cells into the infection sites is a key to enact their protective functions. Importantly, successful early local innate immune response is a critical prerequisite for elicitation of T cell effector functions. Infected tissues often harbor an array of diverse tissue-specific and inflammation-induced chemokines, which guides effector T cell migration and retention. Although many of these chemokines are derived from newly recruited innate immune cells, little is known about how they are presented in the tissue microenvironment and how these innate immune-derived chemotactic signals regulate T cell homing. Here, we show that early recruitment of neutrophils into the influenza-infected trachea is essential for CD8 T cell-mediated immune protection. Both in vitro and in vivo imaging shows that migrating neutrophils leave long-lasting trails from their uropods at the infected trachea. Neutrophil-derived trails contain prominently enriched chemokine CXCL12 and provide both chemotactic and haptotactic cues for efficient CD8 T cell migration and tissue localization. Experiments with granulocyte-specific CXCL12 conditional knockout mice and a CXCR4 antagonist revealed that CXCL12 derived from neutrophil trails is critical for virus-specific CD8 T cell recruitment and anti-viral effector functions. Collectively, these results suggest that early tissue-infiltrating neutrophils deposit chemokine-containing trails, which may function as a long-lasting chemokine depot guiding antigen-specific effector CD8 T cell responses in the influenza-infected tissues. The 2014 Fall Conference of the Korean Association of Immunologists 49 Block Symposium VIII KAI-AKIA Joint Symposium BIOGRAPHY Eui-Cheol Shin Dr. Eui-Cheol Shin graduated Yonsei University College of Medicine, Seoul, Korea in 1996 and received his Ph.D. degree from Yonsei University in 2001 in the major of medical microbiology and immunology. From 2002 to 2007, he researched T cell immune responses in hepatitis C virus infection in NIDDK, National Institutes of Health, Bethesda, MD, USA as a postdoctoral fellow. Since 2007, he has been working as an assistant professor and associate professor in Graduate School of Medical Science and Engineering, KAIST, Daejeon, Korea. His laboratory, Laboratory of Immunology and Infectious Diseases, focuses on the research of T cell immune responses in human viral diseases. [BS VIII-4] Quantitative and Qualitative Changes in Regulatory T Cell Population during Acute Viral Hepatitis Yoon Seok Choi, Jeewon Lee, Eui-Cheol Shin Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, KAIST, Daejeon, Korea Foxp3+ regulatory T (Treg) cells play a major role in maintaining the immune homeostasis. In chronic viral infection, Treg cells have been known to hamper the antiviral T cell responses and reduce immunopathology. In contrast, roles of Treg cells in acute viral infection remain largely unknown. We show here that ex vivo suppressive activity of Treg cells are reduced in patients with acute hepatitis A (AHA). Attenuated suppressive activity of Treg cells was closely correlated with the degree of immune-mediated liver injury. In addition, frequency of circulating Treg cells was also decreased in AHA patients, compared to healthy donors. Furthermore, they contained lower percentage of functionally competent and proliferating population, compared to healthy controls. We also investigated whether functional plasticity of Treg cells is responsible for the impaired function of Treg cells and tissue damage in patients with AHA. We examined the production of a variety of inflammatory cytokines from Treg cells in the context of TCR stimulation. We found that a considerable proportion of Treg cells produced TNF-α following TCR stimulation in patients with AHA. Low level of CD127 expression and demethylated patterns of TSDR (Treg cell-specific demethylated region) in TNF-α+ Treg cell population confirms that TNF-α+ Treg cells have a nature of naturally occurring Treg cells. TNF-α+ Treg cells were enriched in CD45RA−FoxP3lo population, implying their reduced in vivo suppressive activity. Indeed, we observed a close correlation between the frequency of TNF-α+ Treg cells and the clinical severity of liver damage. In conclusion, regulatory roles of Treg cell population are impaired due to both diminished Treg pool size and functional insufficiency of Treg cells in AHA. In addition, TNF-α-producing Treg cells contribute to such functional defectiveness, leading to immune-mediated tissue damage in AHA. 50 The 2014 Fall Conference of the Korean Association of Immunologists The 2014 Fall Conference of the Korean Association of Immunologists Nov. 6(Thu) ~ 7(Fri), 2014 Seoul, Korea Oral Presentation Oral Presentation [OP 1] P-029 Functional Heterogeneity of Influenza Virus-specific CD8+ T Cells 1 2 1 2 2 1 Jihye Kim , Hyeok-Il Kwon , Jeewon Lee , Min-Suk Song , Young Ki Choi , Eui-Cheol Shin * 1 Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology, Daejeon, 2College of Medicine and Medical Research Institute, Chungbuk National University, Cheongju, Korea Tel: 042-350-4266, E-mail: [email protected] CD8+ T cells clear viruses through their two main effector functions: cytolysis and cytokine secretion. However, the factors controlling these antiviral effector activities in vivo at infection sites are poorly understood. In this study, we sug+ gest that IL-15 induces functional heterogeneity of antiviral CD8 effector T cells in the influenza virus-infected lungs. We studied CD8+ T cell responses in a mouse influenza model, using highly virulent (ma81) and its single amino acid substitution (a Iso -to- Thr at position 97) variant, avirulent (ma81-PAI97T) influenza virus strains. We found that CD8+ bright T cells from the lethal influenza (ma81)-infected lungs exerted strong cytotoxicity, evidenced by CD107a , without bright − + IFN-γsecretion (CD107a IFN-γ ). In contrast, conventional effector CD8 T cells with IFN-γsecretion and moderate cytotoxicity (CD107adimIFN-γ+) were observed in the avirulent influenza-infected lungs. Interestingly, the amount of IL-15 was increased in the BAL fluid during the lethal infection, and in vivo anti-IL-15 blocking reduced the frequency of CD107abrightIFN-γ− subset, suggesting that CD107abrightIFN-γ− CD8+ T cells are generated by IL-15. These results indicate the importance of IL-15 on functional heterogeneity of CD8+ T cells to mount appropriate responses in the context of influenza infection. Keywords: Influenza virus, CD8+ T cells, Cytotoxicity, IFN-γ, IL-15 The 2014 Fall Conference of the Korean Association of Immunologists 53 Oral Presentation [OP 2] P-091 Upregulation of PD-1 on Regulatory T Cells Potentiates Their Suppressive Function during Chronic Viral Infection 1 1 1 1 2 Hyo Jin Park *, Joon Seok Park , Yun Hee Jeong , Chan Hee Park , Byoung-Hee Lee , 3 4 5 6 1 Lieping Chen , Jun Chang , Doo Hyun Chung , Inhak Choi , Sang-Jun Ha * 1 Department of Biochemistry, College of Life Science & Biotechnology, Yonsei University, Seoul, 2Division of Biological Resources Coordination, National Institute of Biological Resources, Environmental Research Complex, Incheon, Korea, 3 Department of Immunobiology, Yale University School of Medicine, New Haven, CT 06520, USA, 4 Division of Life & Pharmaceutical Sciences, and Center for Cell Signaling & Drug Discovery Research, Ewha Womans 5 University, Seoul, Department of Pathology, College of Medicine, Seoul National University, Seoul, 6 Department of Microbiology and Immunology, Inje University College of Medicine, Busan, Korea Tel: 02-2123-7584, E-mail: [email protected] Regulatory T (Treg) cells act as terminators in the case of T cell immunity during the acute phase of viral infection; however, their roles in chronic viral infection are not completely understood. Here, we compared the phenotype and function of Treg cells during acute and chronic viral infection using lymphocytic choriomeningitis virus-infected mouse models. Chronic infection, unlike acute infection, led to induction of Treg cells and upregulation of programmed death-1 (PD-1). Treg cells isolated from chronically infected mice displayed greater suppressive capacity for inhibiting T cell proliferation and subsequent cytokine production than those from naïve or acutely infected mice. Suppression was contact-dependent and required PD-1 expression. We found that PD-1 signaling in Treg cells enhanced suppressive function by upregulating IL-10 and granzyme B. These findings establish PD-1 as a mediator of Treg cell suppressive function in the regulation of T cell responses and suggest a role of PD-1 expression on Treg cells, in addition to that on exhausted T cells, during chronic viral infection. Keywords: Programmed death-1, Regulatory T cells, LCMV 54 The 2014 Fall Conference of the Korean Association of Immunologists Oral Presentation [OP 3] P-127 Deficiency of Melanoma Differentiation-associated Protein 5 (MDA5) Results in Exacerbated Chronic Postviral Lung Inflammation 1 1 1 2 Won-keun Kim , Deepika Jain , Melissa D. Sanchez , Cynthia J. Koziol-White , 3 2 2 2 Krystal Matthews , Moyar Q. Ge , Angela Haczku , Reynold A. Panettieri, Jr. , 3 1 Matthew B. Frieman , Carolina B. Lopez * 1 Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, 2Department of Medicine, 3 University of Pennsylvania, Philadelphia, Pennsylvania, Department of Microbiology and Immunology, School of Medicine, University of Maryland at Baltimore, Maryland, USA Tel: 02-920-6354, E-mail: [email protected] Respiratory viral infection can lead to the establishment of chronic lung inflammatory diseases including chronic obstructive pulmonary disease (COPD). Understanding the early innate immune mechanisms that participate in the development of chronic postviral lung disease may be critical for therapeutic intervention. However, little is known of the impact of the innate immune system on the development of long-term lung disease. The melanoma differentiation-associated gene 5 (MDA5) is a germ line encoded pattern recognition receptor (PRR) that plays a role in detecting molecular motifs present in viral oligonucleotides. Sendai virus (SeV) causes bronchiolitis and persistent airway hyperreactivity in mice, providing an accepted experimental model for virus-induced chronic lung disease. Here we show MDA5 deficiency results in a distinct profile of chemokines and cytokines that associated with acute lung neutropenia, enhanced accumulation of alternatively activated macrophages, and sustained lung inflammation upon infection with SeV while viral loads remain unchanged. Remarkably, the biased acute inflammatory response of MDA5 deficiency elicits an enhanced chronic lung inflammation, epithelial cell hyperplasia, airway hyperreactivity, and diminished blood oxygen saturation. Overall, these studies demonstrate that the intracellular viral sensor molecule MDA5 modulates the development of chronic lung inflammation by regulating the early inflammatory response in the lung. Keywords: Respiratory viral infection, Innate immune response, MDA5, Alternatively activated macrophage, Chronic Lung Inflammatory response The 2014 Fall Conference of the Korean Association of Immunologists 55 Oral Presentation [OP 4] P-219 Intravital Imaging of T and B Cell Trafficking Across High Endothelial Venules in Mice Lymph Node 1 1 1 2 2 1,2 Kibaek Choe , Eunjoo Song , Soyeon Ahn , Sukhyun Song , Gou Young Koh , Pilhan Kim * 1 Graduate School of Nanoscience and Technology, 2Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Korea Tel: 042-350-1115, E-mail: [email protected] Lymph node (LN) is a major checkpoint for the circulating lymphocytes to recognize foreign antigens. High endothelial venule (HEV) in LN facilitates an effective recruitment of circulating lymphocytes from the blood. HEV has distinctive cuboidal-shaped endothelial cells and prominent perivascular sheaths consisting of fibro-reticular cells (FRCs). Yet, our understanding about cellular dynamics in transendothelial, intra-perivascular channel and trans-FRC migration in HEV has been relatively limited. In this work, we adapted a custom-design intravital microscopy system to visualize T and B cell migration across HEV in real time in vivo. Actin-DsRed transgenic mice were used to visualize HEV-endothelial cells. FRCs were labeled in vivo by injecting anti-ER-TR7 antibody conjugated with far-red fluorophore into the footpad. T or B cells obtained from actin-GFP mice were adoptively transferred. Multiple steps in transendothelial, intra-perivascular channel and trans-FRC migrations of GFP+ T or B cells were successfully imaged. T and B cells squeezed in between endothelial cells and then migrated along perivascular channel, a narrow space between endothelium and FRCs, for searching a proper site to exit by trans-FRC migration. Interestingly, compared with T cells, B cells spent longer time in passing the perivascular channel although their total moving distances in the perivascular channel were similar. By time-lapse imaging during 2 hours, we found there were preferred exit sites (“exit ramp”) from the perivascular channel for both of T and B cell. To observe whether T and B cell exit through the same exit ramp, we simultaneously transferred DsRed+ T and GFP+ B cells into wild type C57BL/6 mice. Indeed, T and B cells followed each other though the same exit ramp from the perivascular channel. In addition to the exit ramp, we also found that there exists an “entrance ramp” to perivascular channel, a preferred site for transendothelial migration for both of T and B cells. Keywords: Lymph node, High endothelial venule, Lymphocyte, Confocal Microscopy, Intravital Microscopy 56 The 2014 Fall Conference of the Korean Association of Immunologists The 2014 Fall Conference of the Korean Association of Immunologists Nov. 6(Thu) ~ 7(Fri), 2014 Seoul, Korea Poster Presentation Antigen Processing and Presentation (P1-4) Allergy, Hypersensitivity and Autoimmunity (P5-24) Immune Cell Development, Differentiation and Function (P25-49) Immune Mediators, Their Receptors and Signaling (P50-60) Immune Response Regulation and Tolerance (P61-94) Innate Immune Response, Infection and Inflammation (P95-151) Mucosal and Regional Immunity (P152-169) Tumor and Transplantation Immunology (P170-192) Vaccines and Immune Therapeutics (P193-210) Technical Innovations and Others (P211-220) Late Breaking Abstracts (P221-223) Antigen Processing and Presentation P-001 Hepatitis C Virus Suppresses Interferon-Stimulated Production of Immunoproteasomes by the Protein Kinase R Pathway In Soo Oh, Wonseok Kang, Pil Soo Sung, Chung Hwan Cho, Eui-Cheol Shin* Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, KAIST, Daejeon, Korea Tel: 042-350-4236, E-mail: [email protected] P-002 Poster Presentation Measurement of CD8+ and CD4+ T cell frequencies specific for EBV LMP1 and LMP2a using mRNA-transfected DCs Dae-Hee Sohn1, Hyun-Jung Sohn2, Seung-Joo Hyun1, Hyun-Joo Lee2, Shoo-Eon Kim2, Sun-Deok Lee2, Hyun-Il Cho2, Seok-Goo Cho3, Suk-Kyeong Lee4, Tai-Gyu Kim1,2* 1 Department of Microbiology, College of Medicine, The Catholic University of Korea, 2Hematopoietic Stem Cell Bank, College of Medicine, The Catholic University of Korea, 3Hematology, Department of Internal Medi4 cine, College of Medicine, The Catholic University of Korea, Research Institute of Immunobiology, Department of Medical Lifescience, College of Medicine, The Catholic University of Korea Tel: 02-2258-7341, E-mail: [email protected] Background: Hepatitis C Virus (HCV) is known to evade host’s immune responses efficiently. In virus-infected cells, antigen is processed by proteasome complex and presented to CD8+ T cells by major histocompatibility complex (MHC) class I. Interferon (IFN) switches conventional proteasomes to immunoproteasomes, which are more suitable for generation of MHC class I epitopes. Herein, we studied the effect of HCV infection on immunoproteasome production and its mechanism. Methods: JFH-1 (genotype 2a) HCV cell culture system was used for in vitro infection of Huh-7.5 cells. Huh-7.5 cells were treated with type I or II IFNs to induce immunoproteasome production. The expression of immunoproteasome subunits was evaluated by immunoblots and real-time PCR. The expression of protein kinase R (PKR) was silenced with lentiviruses that express small hairpin RNAs, and immunoproteasome induction was examined in PKR silenced cells. Results: Type I or II IFNs induced the expression of immunoproteasome subunits (immunosubunits) such as β1i (LMP2), β2i (MECL-1), and β5i (LMP7). However, IFN-induced immunosubunits expression was attenuated in HCV-infected cells, but not in hepatitis A virus-infected cells. While this attenuation in HCV-infected cells was observed at the protein level, it was not at the mRNA level. This result suggests that IFN-induced immunosubunits expression is hampered during translation in HCV-infected cells. The expression of immunosubunits was restored by PKR silencing in HCV-infected cells, demonstrating that the PKR pathway is responsible for the suppression of immunosubunits expression in HCV-infected cells. Conclusions: HCV suppresses IFN-stimulated expression of immunoproteasome subunits by the PKR pathway. Suppression of immunoproteasome production might contribute to immune evasion of HCV by altering generation of MHC class I epitopes. An EBV-specific cellular immune response is associated with progression of these diseases, some of which have been successfully treated by adoptive T cell therapy. Therefore, many methods have been used to measure EBV-specific cellular immune responses. Previous studies have mainly used autologous EBV-transformed B-lymphoblastoid cell lines (B-LCLs), recombinant viral vectors transfected or peptide pulsed dendritic cells (DCs) as stimulators of CD8+ and CD4+ T lymphocytes. In the present study, we used an interferon-γ (IFN-γ) enzyme-linked immunospot (ELISPOT) assay by using isolated CD8+ and CD4+ T cells stimulated with mRNA-transfected DCs. The frequency of latent membrane protein 1 (LMP1)-specific IFN-γ producing CD4+ T cells was significantly higher than that of LMP2a. In addition, LMP1-specific immune responses were highly correlated with CD8+ and CD4+ T cells. To determine whether there were changes in LMP1- or LMP2a-specific immune responses, subsequent peripheral blood mononuclear cells (PBMCs) samples were analyzed. Significant changes were observed in 5 of the 10 donors examined, and CD4+ T cell responses showed more significant changes than CD8+ T cell responses. CD8+ and CD4+ T cells from EBV-seropositive donors secreted only the Th1 cytokines IFN-γ, TNF-α, and IL-2, while Th2 (IL-4) and Th17 (IL-17a) cytokines were not detected. CD4+ T cells secreted significantly higher cytokine levels than did CD8+ T cells. Analysis of EBV-specific T cell responses using autologous DCs transfected with mRNA might provide a comprehensive tool for monitoring EBV infection and new insights into the pathogenesis of EBV-associated diseases. Keyword: HCV, Immunoproteasome, PKR, Antigen processing Keywords: Epstein-Barr virus, Dendritic cells, mRNA, ELISPOT P-003 Autophagic Protein Atg5 Regulates Tumor Antigen Presentation by Dendritic cells P-004 Dong Sun Oh, Heung Kyu Lee* Lab of Host Defenses, Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Korea Tel: 042-350-4241, E-mail: [email protected] Cancer is still one of the main reasons of death worldwide, although cancer immunotherapy has developed a new treatment for a variety of cancers. Autophagy is known to be important in the presentation of cytosolic and viral antigen on MHC class II. However, the role of autophagic process in tumor antigen presentation in vivo is unclear. Mice with dendritic cell-specific deletion of Atg5, a key autophagic gene, showed the impaired Th1 cell priming, but not cytotoxic T cell response, after apoptotic tumor antigen treatment. The defect of dendritic cells in the absence of Atg5 was the processing and presentation of phagocytosed tumor antigens for MHC class II. In contrast, cross-presentation of peptides on MHC class I in dendritic cells as well as macrophages was not impaired in the absence of Atg5. Thus, our study demonstrated that dendritic cells, but not macrophages, utilize autophagic machinery to process and present apoptotic tumor antigens for MHC class II presentation. Keywords: Tumor, Autophagy, Antigen presentation An MHC Class II Dependent Activation Loop Between Adipose Tissue Macrophages and + CD4 T Cells Controls Obesity-Induced Inflammation Kae Won Cho1,2, David L. Morris1, Jennifer L. 1 1 1 DelProposto , Lynn Geletka , Carey N. Lumeng * 1 Department of Pediatrics and Communicable Diseases, University of Michigan Medical School, Ann Arbor, MI, 48109. USA, 2Soonchunhyang Institute of Medi-bio Science, Soonchunhyang University, Cheonan, Korea Tel: 041-530-4795, E-mail: [email protected] An adaptive immune response triggered by obesity is characterized by the activation of adipose tissue CD4+ T cells by unclear mechanisms. We have examined if interactions between adipose tissue macrophages + (ATMs) and CD4 T cells contribute to adipose tissue metainflammation. Intravital microscopy identifies dynamic antigen dependent interactions between ATMs and T cells in visceral fat. Mice deficient in major histocompatibility complex class II (MHCII) showed protection from diet-induced obesity. Deletion of MHCII expression in macrophages led to an adipose tissue specific decrease in the effector/memory CD4+ T cells, at+ tenuation of CD11c ATM accumulation, and improvement in glucose intolerance by increasing adipose tissue insulin sensitivity. Ablation experiments demonstrated that the maintenance of proliferating conventional T + cells is dependent on signals from CD11c ATMs in obese mice. These studies demonstrate the importance of MHC Class II restricted signals from ATMs that regulate adipose tissue T cell maturation and metainflammation. Keywords: CD4 T lymphocytes, Adipose tissue macrophage, Antigen presenting cells, MHC II The 2014 Fall Conference of the Korean Association of Immunologists 59 Poster Presentation P-005 Allergy, Hypersensitivity and Autoimmunity Blocking Astrocytic Glutamate Carboxypeptidase II Alleviates Glutamate Excitotoxicity and Severity of Experimental Autoimmune Encephalomyelitis P-006 Galectin Isolated from Parasite Inhibits Remission of Experimental Autoimmune Encephalomyelitis by Upregulating Autoantibody 1 1 2 1 Danbee Ha , So Jin Bing , Ginnae Ahn , Jinhee Kim , 1 1 3 Jinhee Cho , Areum Kim , Sangmee Ahn Jo , Youngheun 1 Jee * 1 1 1 2 So Jin Bing , Jinhee Cho , Areum Kim , Sang Kyun Park , 2 1 Hak Sun Yu , Youngheun Jee * 1 2 Department of Veterinary Medicine and Institute for Nuclear Science and 2 Technology , Jeju National University, Jeju, Korea, Laboratory of Veterinary Molecular Pathology and Therapeutics, Division of Animal Science, Graduate School, Tokyo University of Agriculture and Tech3 nology, Fuchu, Japan, Departments of Pharmacy, College of Pharmacy, Dankook University, Cheonan, Korea Tel: 064-754-3374, E-mail: [email protected] Introduction: Experimental autoimmune encephalomyelitis (EAE) is an inflammatory and demyelinating autoimmune disease model of the mouse central nervous system (CNS), and presents clinical similarities to human multiple sclerosis (MS). Although glutamate-induced excitotoxicity has been implicated in the disease progression of MS patient, glutamate carboxipeptidase II (GCPII), an enzyme that produces glutamate and affects the pathogenesis of various neurological disorders, has not received proper attention in MS/EAE. Materials and Methods: EAE was induced in Lewis rats and C57BL/6 and 2-PMPA (10 mg/kg) dissolved in phosphate-buffered saline (PBS) was injected into each mouse intraperitoneally twice a week from the day of inoculation with MOG35-55 until the termination of experiments. Results: When we administered 2-PMPA to EAE mice, markedly attenuated EAE clinical signs were observed along with the significantly inhibited infiltration of inflammatory cells into CNS. Upon antigen restimulation, lack of GCPII led to marked reduction in myelin-reactive T cell responses and T effector cell polarization in periphery. Moreover, GCPII inhibition favorably altered the expression of inducible metabotropic glutamate receptor 1 (mGluR1) on CD4+ T cells. Conclusion: These data suggest previously unreported interaction between astrocytic GCPII expression and critical EAE-associated features including neuroinflammation and highlight mGluR1 as one of key modulator of glutamate-induced T cell responses during EAE. 1 College of Veterinary Medicine, Jeju National University, Jeju, Department of Parasitology, School of Medicine, Pusan National University, Yangsan, Korea Tel: 064-754-3374, E-mail: [email protected] Recently, parasite infections or parasite-derived products have been suggested as a therapeutic strategy with suppression of immunopathology involves the induction of regulatory T cells or/and Th2 responses. Among multiple potential immunomodulatory protein families in parasites, galectins is particularly known to multifunctional immune mediators in the mammalian immune system. Although many instances of mammalian galectins acting to suppress immunological reactions, there is still insufficient information whether galectin expressed by parasite also perform similar functions as mammalian galectins or have own distinct function associated with their virulence. In a recent study, researchers reported that constructed recombinant galectin (rTl-gal) isolated from an adult worm of the gastrointestinal nematode parasite Toxascaris leonina attenuated clinical symptoms of inflammatory bowel disease in mice treated with dextran sulfate sodium (DSS). Noting that the role of rTl-gal in the inflammatory disease, we attempted to confirm the effect of parasite via its rTl-gal on other neuronal autoimmune disease using experimental autoimmune encephalomyelitis (EAE), a mouse inflammatory and demyelinating autoimmune disease model of human multiple sclerosis (MS). In this model, rTl-gal treated EAE mice did not fail to recover after the peak of the disease, leading to persistent CNS damages such as demyelination, gliosis and axonal damages. Further, rTl-gal treated EAE mice markedly increased the number of CD45R/B220+ B cells in both infiltrated inflammation and periphery along with increase the production of proinflammatory cytokines and pathogenic autoantibody. Our results suggest that galectin isolated from a gastrointestinal parasite can deliver a harmful effect to EAE via enhancing B cell function contrary to its beneficial effect on inflammatory bowel disease. This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (NRF-2012R1A2A2A01011645) This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (NRF-2012R1A2A2A01011645 and NRF-2012-Global Ph.D. Fellowship Program). Keywords: Glutamate carboxypeptidase II, Glutamate excitotoxicity, Experimental autoimmune encephalomyelitis (EAE) Keywords: Parasite galectin, Toxascaris leonina, Experimental autoimmune encephalomyelitis (EAE) P-007 Synergistic Effects of Silver Nanoparticles in Atopic Dermatitis Mice Model Seung Jae Kim1, Han Goo Kang1,2, In-Hong Choi1,2,3* 1 Department of Microbiology, 2Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine and 3Institute for Immunology and Immunological Disease Tel: 02-2228-1821, E-mail: [email protected] Atopic dermatitis (AD) is a chronic and relapsing allergic skin disease that accompanies itching and eczema. Mast cell degranulation, major characteristics in AD, is occurred when IgE links with antigens and then, various inflammatory cytokines are released. Recent studies suggest that silver nanoparticles (AgNPs) have cytotoxic effects in mammalian cells. Moreover, allergic effects were elevated by inhaled AgNPs in the allergen-provoked mice model. In this study, we investigated synergistic effects of AgNPs in AD mice model. The extent of AD was evaluated with serum IgE level, atopic dermatitis scores, ear thickness and histopathological observation of mast cell infiltration in dermis. Treatment of 5 nm AgNPs enhanced AD features in a dose-dependent manner but 100 nm AgNPs did not. Serum IgE level was elevated in 5 nm AgNPs group compared with AD control group or 100 nm AgNP-treated group. Atopic dermatitis scores, ear thickness and mast cell infiltration were also increased in 5 nm AgNP-treated group but not in AD control group or 100 nm AgNP-treated group. AD was not induced in mice treated with 5 nm AgNPs alone. Our studies demonstrate that 5 nm AgNPs do not induce AD per se but show synergistic effects in AD mice model. Keywords: Atopic dermatitis, Allergy, Silver nanoparticles, Nanometerials P-008 Preferential Induction of the Novel B7 Family Member, 4Ig B7-H3, on Synovial Monocytes in Rheumatoid Arthritis (RA) Bo Ruem Yoon1, Yeon-Ho Chung1, Su-Jin Yoo2, Seong 2 1,3 Wook Kang *, Won-Woo Lee * 1 Department of Microbiology and Immunology, Seoul National University 2 College of Medicine, Department of Internal Medicine, Chungnam National University School of Medicine, 3Department of Biomedical Sciences, Seoul National University College of Medicine Tel: 02-740-8545, E-mail: [email protected] B7-H3, a newly identified B7 family member, has functional duality as a T-cell costimulator and coinhibitor that fine-tunes T-cell mediated immune responses. Given that B7-H3 expression on human monocytes and dendritic cells is enhanced by inflammatory cytokines, a potential inmmunoregulatory role for this molecule at sites of inflammation has been suggested. Further, monocytes play crucial roles in the pathophysiology of various inflammatory disorders including autoimmune diseases; however, the immunological role of B7-H3 in rheumatoid arthritis (RA) has not been defined. Thus, we aimed to investigate the possible roles of monocyte B7-H3 in the pathogenesis of RA. Here we found that synovial monocytes, but not peripheral monocytes, in RA patients predominantly express surface B7-H3. The 4Ig isoform of B7-H3 is exclusively induced on the cell surface, whereas the 2Ig B7-H3 isoform is constitutively expressed in the intracytoplasmic region of both peripheral and synovial monocytes. Further, B7-H3 knockdown experiments provide evidence that surface B7-H3 has an inhibitory effect on interferon-gamma (IFNγ) production in CD4 memory cells. Moreover, the expression level of 4Ig B7-H3 on synovial monocytes inversely correlates with clinical parameters such as C-reactive protein (CRP) levels, erythrocyte sedimentation rates (ESR), and Disease Activity Scores in 28 joints (DAS28). Our findings demonstrate that activation-induced B7-H3 expression on synovial monocytes has the potential to inhibit Th1-mediated immune responses and immunomodulatory roles thereby affecting the pathogenesis of RA. Keywords: B7-H3, Rheumatoid arthritis (RA), Monocyte, Coinhibitory molecule, Synovial fluid 60 The 2014 Fall Conference of the Korean Association of Immunologists Allergy, Hypersensitivity and Autoimmunity P-009 Identification and Validation of Novel Autoantibodies in Type 1 Diabetic Patients 1 2,3 2,4 Eunhee G. Kim , Bo Kyung Koo , Kyong Soo Park , 4 1 Eugene C. Yi , Kristine M. Kim * 1 Department of Systems Immunology, College of Biomedical Science, and Institute of Bioscience and Biotechnology, Kangwon National University, Chuncheon, 2Department of Internal Medicine, Seoul National University College of Medicine, Seoul, 3Department of Internal Medicine, Boramae 4 Medical Center, Seoul, Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, and College of Medicine or College of Pharmacy, Seoul National University, Seoul, Korea Tel: 033-250-8382, E-mail: [email protected] Type 1 diabetes (T1DM) is autoimmunedisease that results from immune-mediated pancreatic beta cell destruction. T1DM dependent autoantibodies (AAbs) can facilitate diagnostic and therapeutic for T1DM patients. We have identified two novel AAb candidates, anti-EEF1A1 and anti-UBE2L3,from serum samples of T1DM patients, and compared to the serum samples of Type 2 diabetic (T2DM) patients and healthy control subjects with normal glucose tolerance (NGT) using protein microarrays. These novel AAbs were further validated by immunofluorescence staining of pancreas and by ELISA in two independent test cohorts: 1) 95 adults with T1DM, 49 with T2DM, 11 with latent autoimmune diabetesin adults (LADA), 20 with Graves disease, and 66 with NGT; and 2) age-matched 101 T1DM and 150 healthy subjects. The titer of anti-EEF1A1 and anti-UBE2L3 antibodies was significantly higher in T1DM patients (P < 0.001 for both AAbs) than other sample groups. Additionally, prevalence of anti-EEF1A1 and anti- UBE2L3 antibodies was 15.8% and 10.9% in age-matched T1DM patients, respectively. Our method demonstrates identification and validation of AAbs that can provide a new insight into diagnosis and classification of T1DM. Keywords: Autoantibody, Type 1 diabetes (T1DM). Autoimmune disease P-010 Poster Presentation Caffeic Acid Phenethyl Ester Lessens Disease Symptoms in Experimental Autoimmune Uveoretinitis Mouse Model Jae-Hyeog Choi1,2, Kug-Hwan Roh1,2, Hana Oh2,3, Sol-Ji Park1,2, Sung-Min Ha1,2, Mi-Seon Kang4, Ji-Hyun Lee2, Hyun-Keun Song1,2, Jae-Wook Yang2,3*, SaeGwang 1,2 Park * 1 Department of Microbiology and Immunology, College of Medicine, Inje 2 University, Ocular Neovascular disease Research Center, Busan Paik Hospital, 3Department of Ophthalmology, Busan Paik Hospital, College of Medicine, Inje University, 4Department of Pathology, Busan Paik Hospital, College of Medicine, Inje University, Busan, Korea Tel: 051-890-8702, E-mail: [email protected] Experimental autoimmune uveoretinitis (EAU) is an autoimmune disease that models human uveitis. Caffeic acid phenethyl ester (CAPE) is a phenolic compound isolated from propolis and possesses anti-inflammatory and immunomodulatory properties. CAPE demonstrates therapeutic potential in several animal disease models through its ability to inhibit NFκB activity. To evaluate these therapeutic effects in EAU, we administered CAPE in a model of EAU that develops after immunization with interphotoreceptor retinal-binding protein (IRBP) in B10.RIII and C57BL/6 mice. Significantly, we were able to demonstrate that CAPE lessened the severity of EAU symptoms in both mouse strains. Notably, treated mice exhibited a decrease in the ocular infiltration of immune cell populations into the retina, reduced TNF-α, IL-6, and IFN-γserum levels, as well as inhibited TNF-α mRNA expression in retinal tissues. Although CAPE failed to inhibit IRBP-specific T cell proliferation, it was sufficient to suppress cytokine production. In addition, retinal tissues isolated from CAPE-treated EAU mice revealed a decrease in NF-κB p65 and phospho-IκBα. The data identify CAPE as a potential therapeutic agent for autoimmune uveitis that acts by inhibiting cellular infiltration into the retina, reducing the levels of pro-inflammatory cytokines and blocking NF-κB pathway activation. Keywords: Experimental autoimmune uveoretinitis, Caffeic acid phenethyl ester, Pro-inflammatory cytokine, Nuclear factor-kappa B P-011 Identification of Autoantibodies to Aquaporin-5 in Sera from Patients with Sjögren’s P-012 Effect of Vanillic Acid on the Allergic Airway Inflammation in Mouse Model Jehan Alam, Yun Sik Choi, Youngnim Choi* Ji-Hyun Choi, Young-Joo Yi*, Sang-Myeong Lee* Department of Microbiology and Immunology School of Dentistry, Seoul National University, Seoul, Korea Tel: 02-740-8643, E-mail: [email protected] Division of Biotechnology, Chonbuk National University, Iksan, Korea Tel: 063-850-0835, E-mail: [email protected] Sjögren’s syndrome (SS) is an autoimmune disorder that primarily targets the salivary and lacrimal glands, leading to dryness of the mouth and eye. Anti-Ro/SSA and/or anti-La/SSB autoantibodies are the hall mark of disease diagnosis and present in 60-70% of the patient. A number of autoantibodies have been identified in SS, including anti-salivary gland protein1 (SP-1), anti-carbonic anhydrase 6 (CA6), anti-parotid secretory protein (PSP), anti-muscarinic receptor 3, anti-tissue kallikrein, and anti-α-fodrin antibodies. Although anti-muscarinic receptor 3 autoantibodies have been shown to inhibit the function of acinar cells, the pathophysiology of exocrine dysfunction in SS is not fully understood. Aquaporin-5 (AQP5) is a water-channel protein expressed at the acinar cells of the lacrimal and salivary glands, and plays a critical role in tear and saliva secretion. We hypothesized that SS patients may have autoantibodies to AQP5 in sera. To test our hypothesis, frozen section of mouse submandibular salivary gland were dual stained with anti-AQP5 and control/SS patient sera. AQP5 expressed in mucous acini, serous acini, and ductal areas showed strong co-localization with the signals stained with SS patient IgG but not with those by control sera. The presence of autoantibodies to AQP5 in SS sera was further confirmed using Chinese Hamster Ovary (CHO) cells that overexpressed a human AQP5-EGFP fusion protein. Sera from SS patients but not from control subjects stained selectively the AQP5-EGFP but not EGFP expressed in CHO cells. In addition, immunoprecipitation of the lysates of HEK-293 cells overexpressing AQP5-EGFP with SS sera selectively pulled down AQP5-EGFP. In conclusion we present for the first time that SS sera contain anti-AQP5 autoantibodies, which have a potential to be a useful disease biomarker. Asthma is one of well-known inflammatory diseases, which is growing as the public health issue due to environmental pollutions. Vanillic acid (VA) is an oxidized form of vanillin that is used as a flavoring ingredient. Recently, VA has been shown to prevent development of inflammatory diseases such as acute liver injury and dextran sulfate sodium (DSS)-induced ulcerative colitis. However, the effects of VA have not been reported in the allergic airway inflammation. In the present study, we investigated the anti-asthmatic effects of VA using mouse model of asthma. Female BALB/C mice were immunized with ovalbumin (OVA) and were exposed to inhalation with 3% OVA for 30 min on days 28 to 30. VA (1 mg/kg/day) was administered to mice by intraperitoneal injection on days 25 to 30. We found that VA treatment significantly decreased lymphocytes infiltration into airway and the production of Th2 cytokines, such as IL-5 and IL-13 in bronchoalveolar lavage fluids (BALF). Moreover, significantly lower level of IgE was also detected in the serum of mice with VA-injection than controls. In quantitative RT-PCR, VA remarkably suppressed mRNA expressions of IL-4, IL-5, IL-13, and IFN-γ. The lower levels of IL-17 and RORγt mRNA were also detected in mesenteric lymph nodes (MLN) from mice with VA. In conclusion, VA sufficiently reduced the recruitment of immune cells and the production of Th2 cytokines, suggesting that could cure allergic asthma as a potential therapeutic agent. Keywords: Inflammation, Asthma, BALF, Vanillic acid, Mouse Keywords: Autoantibodies, AQP5 Aquaporin 5, Sjögren's syndrome, Dry mouth The 2014 Fall Conference of the Korean Association of Immunologists 61 Poster Presentation P-013 Allergy, Hypersensitivity and Autoimmunity Two Distinct Subpopulations of γδ T Cells Secrete IL-17A by TCR and Cytokine Stimulation in Psoriatic Inflammation Jong Hoon Kim1,2, Young Joon Choi1, Mi-Young Song3, Chae Yeon Ban1, Song-Ee Kim2, Jihye Kim1, Tae-Gyun Kim4, Hyoung-Pyo Kim4, Young-Chul Sung3, Soo-Chan 2 1 Kim *, Eui-Cheol Shin * 1 Laboratory of Immunology and Infectious Diseases, Graduate School of 2 Medical Science and Engineering, KAIST, Daejeon, Department of Dermatology and Cutaneous Biology Research Institute, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, 3Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology, Pohang, 4Department of Environmental Medical Biology, Institute of Tropical Medicine, Yonsei University College of Medicine, Seoul, Korea Tel: 042-350-4236, E-mail: [email protected] Psoriasis is IL-17-mediated chronic inflammatory skin disease and IL-17-producing γδT (γδT17) cells have a critical role in psoriatic inflammation. Among γδT17 cells, Vγ4+ and Vγ6+ cells were pathogenic to induce Imiquimod (IMQ)-induced psoriatic inflammation, but it remains unclear which stimulus activates two subpopulations of γδT cells to produce IL-17. Despite of TCR hypo-responsiveness of γδT17 cells in physiologic condition, we observed that TCR sensitivity of the cells is highly increased after IMQ application. Here, we demonstrate that TCR recognition and cytokines including IL-23 and IL-1β differentially activate two types of γδT17 cells in + IMQ-induced psoriatic inflammation. Vγ4 cells are more sensitive to secrete IL-17A under IL-1β and IL-23 stimulation than Vγ1−Vγ4− cells, mainly containing Vγ6+ subtype, whereas Vγ1−Vγ4− cells preferentially secrete + IL-17A upon TCR stimulation than do Vγ4 cells. In addition, we found up-regulation of PD-1, ICOS and down-regulation of CD127 on the surface of activated γδT17 cells in this model. Of two subsets, the levels of PD-1 and − − ICOS are higher in a Vγ1 Vγ4 subset, while CD127 is more highly expressed in a Vγ4+ subset. These studies dissect distinctively activated two subpopulations of γδ T cells depending on the types of stimuli in psoriatic inflammation. P-014 Hydroxysafflor Yellow A Attenuates Allergic Airway Inflammation by Suppressing the Activity of Nuclear Factor-kappa B in Asthmatic Mice 1 2 1 Qianfei Xue , Guang Hai Yan , Yun Ho Choi * 1 Department of Anatomy, Medical School, Institute for Medical Sciences, Chonbuk National University, Jeonju, Korea, 2Department of Anatomy and Histology and Embryology, Yanbian University School of Basic Medical Sciences, Jilin, China Tel: 063-270-3082, E-mail: [email protected] Safflower is the dry flower of Carthamus tinctorius L. and naturally distributed around the world. Hydroxysafflor yellow A (HSYA), the main active ingredient in safflower, is shown to reduce several inflammatory responses mediated by various stimuli or cytokines. Thus, the objective of our study is to determine whether the HSYA attenuates inflammatory response in an ovalbumin (OVA)-induced asthma model. OVA-sensitized/challenged mice represented the inflammatory cell infiltration in lung tissues as well as airway hyperresponsiveness (AHR) to inhaled methacholine. In addition, these mice had an increased amount of eosinophils, T-helper2 type cytokines [interleukin (IL)-4, IL-5, and IL-13], eotaxin, and adhesion molecules in their bronchoalveolar lavage fluids. However, the administration of HSYA before the OVA challenge resulted in significant inhibition of these asthmatic reactions. Moreover, OVA significantly increased the transcriptional activity of nuclear factor-kappa B (NF-κB) in lung tissues, whereas HSYA markedly suppressed NF-κB translocation. These findings indicate that HSYA protects against OVA-induced airway inflammation and AHR, at least in part via downregulation of NF-κB activity. Our data support the utility of HSYA as a potential medicine for the treatment of asthma. Keywords: Hydroxysafflor yellow A, Airway inflammation, T-helper2 type cytokine, and NF-κB Keywords: γδ T cell, IL-17, Psoriasis, Imiquimod, TCR responsiveness P-015 The Effect of Salidroside on Cytokines and Airway Inflammation of Asthma Induced by Ovalbumin in Mice by Regulating NF-κB and p38 MAPK Activities 1 2 1 Qianfei Xue , Guang Hai Yan , Yun Ho Choi * 1 Department of Anatomy, Medical School, Institute for Medical Sciences, 2 Chonbuk National University, Jeonju, Korea, Department of Anatomy and Histology and Embryology, Yanbian University School of Basic Medical Sciences, Jilin, China Tel: 063-270-3082, E-mail: [email protected] Salidroside is a biologically active ingredient of Rhodiola rosea, which has several interesting biological properties, including anti-oxidant and anti-inflammatory; however, its anti-allergic effects are poorly understood. The objective of this study is to determine whether salidroside attenuates inflammatory response in an ovalbumin (OVA)-induced asthma model. OVA-sensitized/challenged mice show airway hyperresponsiveness (AHR) to inhaled methacholine, and have an increased amount of T-helper2 type cytokines (interleukin (IL)-4, IL-5, and IL-13) and eosinophils in their bronchoalveolar lavage fluids and lung tissues. However, three successive intraperitoneal administrations of salidroside before the last OVA challenge result in significant inhibition of these asthmatic reactions. Moreover, OVA significantly increases the activation of nuclear factor-kappa B (NF-κB) and p38 mitogen-activated protein kinase (MAPK) in lung tissues, whereas salidroside markedly suppresses NF-κ B translocation, and reduces phosphorylation of p38 MAPK. Furthermore, salidroside attenuates the expression of intercellular adhesion molecule 1 and IL-6 through modulating the activities of p38 MAPK and NFκB in the BEAS-2B cells stimulated by proinflammatory cytokines. These findings indicate that salidroside protects against OVA-induced airway inflammation and AHR, at least in part via downregulation of NFκB and p38 MAPK activities. Our data support the utility of salidroside as a potential medicine for the treatment of asthma. Keywords: Salidroside, Airway inflammation, NF-κB, p38 62 P-016 Human Umbilical Cord Blood Mesenchymal Stem Cell-derived PGE2 and TGF-β1 Alleviate Atopic Dermatitis by Reducing Mast Cell Degranulation 1,3 1,2 1 Hyung-Sik Kim , Jun-Won Yun , Tae-Hoon Shin , 3 1,3 Kwang-Won Seo , Kyung-Sun Kang * 1 Adult Stem Cell Research Center, College of Veterinary Medicine, Seoul National University, Seoul, 2Department of Experimental Animal Research, Biomedical Research Institute, Seoul National University Hospital, Seoul, 3 Institute for Stem Cell and Regenerative Medicine in Kangstem Biotech, #406 Biotechnology Incubating Center, Seoul National University, Seoul, Korea Tel: 02-880-1246, E-mail: [email protected] Mesenchymal stem cell (MSC) is a promising tool for the therapy of immune disorders. However, their efficacy and mechanisms in treating allergic skin disorders are less verified. We sought to investigate the therapeutic efficacy of human umbilical cord blood-derived MSCs (hUCB-MSCs) against murine atopic dermatitis (AD) and to explore distinct mechanisms that regulate their efficacy. AD was induced in mice by the topical application of Dermatophagoides farinae. naïve or activated-hUCB-MSCs were administered to mice, and clinical severity was determined. The subcutaneous administration of nucleotide-binding oligomerization domain2 (NOD2)-activated hUCB-MSCs exhibited prominent protective effects against AD, and suppressed the infiltration and degranulation of mast cells (MCs). A β-hexosaminidase assay was performed to evaluate the effect of hUCB-MSCs on MC degranulation. NOD2-activated MSCs reduced the MC degranulation via NOD2-COX2 signaling. In contrast to bone marrow-derived MSCs, hUCB-MSCs exerted a cell-to-cell contact-independent suppressive effect on MC degranulation through the higher production of prostaglandin E2 (PGE2). Additionally, TGFβ1 production from hUCB-MSCs in response to IL-4 contributed to the attenuation of MC degranulation by down-regulating FCεRI expression in MCs. In conclusion, the subcutaneous application of NOD2-activated hUCB-MSCs can efficiently ameliorate AD, and MSC-derived PGE2 and TGF-β1 are required for the inhibition of MC degranulation. Keywords: Mesenchymal stem cells, Atopic dermatitis, Mast cell degranulation, Immunomodulation, NOD2 The 2014 Fall Conference of the Korean Association of Immunologists Allergy, Hypersensitivity and Autoimmunity P-017 Expansion of CD80+ Autoreactive Marginal Zone B Cells Reactive to Type II Collagen in the Collagen-Induced Arthritis Chanho Park, Joo Hee Lee, In Seong Kho, Tae Jin Kim* Department of Immunobiology and Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, Suwon, Korea Tel: 031-299-6161, E-mail: [email protected] Collagen-induced arthritis (CIA), an experimental model of rheumatoid arthritis, is an autoimmune disease induced by immunization with heterologous type II collagen. The CIA induction is dependent on both T and B cells and the generation of autoreactive IgG antibody against type II collagen is essential for the CIA pathogenesis. The CIA is induced in the DBA/1J mice, but not in C57BL/6 mice. So far it is not well known why the DBA/1J strain is more susceptible to CIA than the C57BL/6 strain. + Here we demonstrate the abundance of CD80 marginal zone (MZ) B cells in the DBA/1J mice, which harbor autoreactive B cells capable of producing IgM antibodies reactive to type II collagen. In contrast, MZ B cells from C57BL/6 mice did not express CD80. Furthermore, the anti-type II collagen antibody secretion was found only in theCD80+ MZ B − cells from DBA/1J mice, but not in CD80 MZ B cells in the ELISA and ELISPOT analyses. Upon the immunization with heterologous type II collagen, anti-type II collagen Ig G responses were distinctively revealed in the DBA/1J mice, but not in C57BL/6 mice. We also observed the grad+ + low high ual expansion of MZ B cells that were B220 , CD21 , CD23 CD38 , IgMhigh, and especially CD80+ with the disease progression in the DBA/1J + CIA model. Based on our results, we suggest that the autoreactive CD80 MZ B cells, which were uniquely present in the DBA/1J mice, may be responsible for the susceptibility of the strain to the CIA. P-018 Poster Presentation A Novel Biguanide Derivative MD-6 Inhibits IL-17 Production and Attenuates Murine Autoimmune Arthritis Kihwa Lee, Youngjun Park, Minji Kim, Sun-A Im, Sukgil Song, Chong-Kil Lee* Department of Pharmacy, Chungbuk National University, Cheongju, Korea Tel: 043-261-2826, E-mail: [email protected] Metformin is one of the most used drugs as the first line agent to treat type 2 diabetes mellitus. Recent reports showed that metformin mitigates murine autoimmune arthritis and reduces Th 17 cell differentiation. In this study, the anti-autoimmune arthritis effect of MD-6, a derivative of metformin, were investigated. MD-6 inhibited the proliferation of mouse splenocytes that were stimulated with IL-2. MD-6 also inhibited the production of proinflammatory cytokines from splenocytes that were stimulated with anti-CD3 and anti-CD28 antibodies. MD-6 exerted 1000-fold strong anti-infiammatory activity in vitro compared with metformin. To evaluate the effect of MD-6 on collagen antibody-induced arthritis (CAIA), mice were administered intraperitoneally with either MD-6 or metformin on a daily basis, starting just after the injection of the anti-type Ⅱ collagen antibody, for four weeks. MD-6 efficiently attenuated the severity of CAIA and decreased the serum level of IL-17. MD-6 exerted 10-fold strong anti-arthritis activity in vivo compared with metformin. These findings reveal that MD-6 has a therapeutic value for the treatment of rheumatoid arthritis and other inflammatory diseases. Keywords: Metformin, MD-6, Th17 cell, Autoimmune arthritis Keywords: Rheumatoid arthritis, Collagen-induced arthritis, Marginal zone B cells P-019 Porphyromonas gingivalis HSP60 Peptides Have Distinct Roles in the Development of Atherosclerosis Euikyong Jeong1, Gil Sun Cha2, Sung-Jo Kim2, Ho Sung 1 2 Kang , Jeomil Choi * 1 Department of Molecular Biology, College of Natural Sciences, Pusan 2 National University, Department of Periodontology, School of Dentistry, Pusan National University, Dental Research Institute Tel: 055-360-5200, E-mail: [email protected] Background: Different epitope peptides of bacterial heat shock proteins may function as effector or regulatory molecules in autoimmune responses in infection-triggered atherosclerosis. Study Aim: We investigated the mechanisms for the distinct roles of two epitope peptides from P. gingivalis heat shock protein 60 (HSP60) in atherogenesis with regard to peptide-specific T cell polarization relevant to 1) phenotype and cytokine profiles, 2) expression of transcription factors, and 3) role of antigen presenting dendritic cell subsets. Materials and Methods: Apolipoprotein E-knockout (ApoE KO) mice were immunized with peptide 14 or peptide 19 from P. gingivalis HSP60 prior to induction of atherosclerosis by infection with P. gingivalis plus a Western diet. Results: Significant reductions in plaque/lipid droplet area and plasma cholesterol levels were observed in mice immunized with peptide 14, whereas the opposite phenomenon + was evident in mice immunized with peptide 19. CD4 T-cells polarized to the regulatory T-cell type in peptide 14-immunized group, whereas they polarized to the Th1 cells in peptide 19-immunized group; this observation was supported by the cytokine profiles characteristic to each T-cell phenotype. Significantly higher expression of Nr4a1 and Nr4a2 mRNA, transcriptional factors for regulatory T-cell type, were observed in peptide 14-immunized group. In contrast, the expression level of IFN-γ and T-bet mRNA, signaling molecules for Th1 cells, was higher in peptide 19-immunized group than in + PBS-immunized group. In non-immunized wild mice, BMDC-derived CD11c dendritic cells have shown to stimulate Tregs significantly in antigen-nonspecific manner. However, each peptide antigen demonstrated a unique mode of preferential adoption of dendritic cell subsets. Conclusion: Peptide 14 or peptide 19 from P. gingivalis HSP60, respectively, may play either an anti- or pro-atherogenic role in the ApoE KO mouse model of infection-triggered atherosclerosis through distinct mechanisms operating in the polarization of T cells. P-020 IL-7 Plays a Crucial Role for Allergic Th2 Memory Cells to Maintain Their Homeostasis in Murine Allergic Asthma Model Seung-min Yeon, Yong Woo Jung* Department of Pharmacy, Korea University, Sejong, Korea Tel: 041-860-1618, E-mail: [email protected] Memory T cells efficiently protect host from harmful pathogens because of their longevity, robust response and homeostatic proliferation. Although memory T cells are well characterized in response to infectious agents, allergen-specific CD4 memory T cells have been poorly studied. Since the CD4 Type 2 helper T (Th2) cells play a significant role in the development and the maintenance of allergic asthma, we hypothesized that allergic memory Th2 cells are present and specialized microenvironments provide key factors for the homeostasis of these cells. We found memory Th2 cells mainly present in the lungs and airways in a murine asthma model. These cells showed memory characteristics with their longevity, rapid secondary response and homeostatic proliferation. In addition, the expression of Interleukin 7 (IL-7) increased after allergen challenges and the blockade of this cytokine severely reduced their half-life. By the contrary, the expression of Interleukin 25 (IL-25) mRNA decreased during the contraction phase. In conclusion, allergic memory Th2 cells are present and IL-7 plays a crucial role in maintaining their homeostasis. Keywords: Allergic asthma, Th2 memory cell, IL-7, Homeostatic proliferation This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (NRF-2011-0015501) and 2014 Clinical Research Grant, Pusan National University Dental Hospital. Keywords: Atherosclerosis, Periodontitis, Autoimmunity, Porphyromonas gingivalis, Heat shock protein, Regulatory T cell, Transcriptional factors The 2014 Fall Conference of the Korean Association of Immunologists 63 Poster Presentation P-021 Allergy, Hypersensitivity and Autoimmunity Inhibitory Effect of MPG416 on Allergic Response Hae-Sim Cha, Tack-Joong Kim* Division of Biological Science and Technology, College of Science and Technology, Yonsei University, Wonju, Korea Tel: 033-760-2242, E-mail: [email protected] Mast cells are recognized as the key mediators of IgE-mediated allergic diseases including asthma, rhinitis, and atopic dermatitis (AD). The purpose of this study was to evaluate the anti-allergic activity of MPG416 using activated mast cells and an AD-like ICR mouse model. We observed that MPG416 decreased mast cell degranulation, decreased the phosphorylation of spleen tyrosine kinase and down-stream signaling molecules. MPG416 suppressed the production of intracellular ROS and the expression of interleukin 4 mRNA. In addition, MPG416 effectively suppressed the passive cutaneous anaphylaxis (PCA) reaction in vivo. In a 2,4-dinitrochlorobenzene-induced AD-like mouse model, MPG416 reduced the serum IgE level, scratching behavior, skin severity scores, and mRNA expression of inflammatory cytokines. These results indicate that MPG416 inhibits the allergic response by reducing mast cell activation and may have clinical potential as an anti-allergic agent in disorders such as atopic dermatitis. Keywords: Atopic dermatitis, MPG416, Passive cutaneous anaphylaxis P-022 Glutamine Efficiently Prevents Biphasic Anaphylaxis via MKP-1-dependent cPLA2 Deactivation Hae-Kyoung Kim, Jae Seok Jeong, Yoo-Dong Lee, Yu-na Im, Hern-ku Lee* Department of Immunology, Chonbuk National University Medical School, Jeonju, Korea Tel: 063-270-3069, E-mail: [email protected] It has been reported that cytosolic phospholipase A2 (cPLA2) is critical for the development of late anaphylactic reaction (LAR). In this study, the therapeutic efficacy of the non-essential amino acid, L-glutamine (Gln) was investigated, given that Gln functions as a potent cPLA2 inhibitor. cPLA2 inhibitors, pyrrophenone and cPLA2 siRNA as well as, as cPLA2 metabolites, leukotriene (LT)B4 and platelet-activating-factor (PAF) prevented LAR. Gln efficiently prevented LAR, as well as lung levels of LTB4 when Gln was administered up to 3 h after challenge injection. Gln injection resulted in inhibition of challenge-induced cPLA2 phosphorylation as well as induction of MAPK phosphatase (MKP)-1. MKP-1 siRNA abrogated the Gln-mediated i) prevention of LAR, 2) deactivation of cPLA2, and iii) induction of MKP-1. ERK inhibitor, PD98059 abolished Gln-mediated MKP-1 induction. These results suggest that Gln inhibits LAR via deactivation of cPLA2, resulting in inhibition of its metabolites production such as LTB4 and PAF, through the induction of MKP-1 in an ERK-dependent manner. Keywords: L-Glutamine, Anaphylaxis, Late anaphylactic reaction, cPLA2, cytosolic phospholipase A2, Leukotriene B4 P-023 Sanroque Mice Develop Sjögren Syndrome-Like Disease with Age Eun Kyeong Jang1, Yeon Kyung Oh1, Kiseok Jang2, Jeehee Youn1* 1 Laboratory of Autoimmunology, Department of Biomedical Science, Department of Anatomy & Cell Biology, 2Department of Pathology, Hanyang University, Seoul, Korea Tel: 02-2220-0604, E-mail: [email protected] Sjögren syndrome (SS) is a chronic autoimmune disorder that affects mainly salivary and lacrimal glands, leading to dry month and eyes. Along with the deficiency of animal models for human SS, its etiopathogenesis largely remains to be explored. Here we examined whether Sanroque mice have a potential to act as a model for SS. Sanroque mice bear an M199R mutation of Roquin (Rc3h1) which function as an ICOS repressor. Sanroque homozygous females at the age of 10-20 weeks develop follicular helper T cells and anti-dsDNA antibodies at the higher levels than their wild-type littermates, but other inflammatory symptoms were not accompanied at this age. However, the mice at the age of more than 40 weeks spontaneously developed signs of SS; Saliva flow was significantly reduced, and salivary glands were severely infiltrated by lymphoid cells including plasma cells. Importantly, most plasma cells residing in the inflamed salivary glands were long-lived plasma cells. Glomerulonephritis in the kidney was also evident in the aged Sanroque mice. Thus, our results suggest that chronically dysregulated activity of follicular helper cells is sufficient to precipitate symptoms of SS and propose Sanoque mice as a model of human SS. Supported by the Korean Health Technology R&D Project, Ministry of Health & Welfare, Korea (HI13C0016). P-024 Early Growth Response-1 Promotes Differentiation of Plasma Cells Yeon Kyung Oh, Eun Kyeong Jang, Jeehee Youn* Laboratory of Autoimmunology, Department of Biomedical Science, Department of Anatomy & Cell Biology, Hanyang University, Seoul, Korea Tel: 02-2220-0604, E-mail: [email protected] Early growth response (EGR)-1 is a Cys2-His2-type zinc-finger transcription factor. Egr-1 has been known to induce apoptosis and growth in immature and mature B cells, respectively, but its role in the differentiation of B cells to plasma cells remains obscure. Here we examined whether Egr-1 is important for B cells to commit to plasma cell fates using Egr-1 knockout (KO) mice. naïve B cells from Egr-1 wild-type (WT) or KO mice were stimulated with LPS and IL-4, and the resulting cells were assayed by FACS. Cell viability and apoptosis were not different between WT and KO cells, but the differentiation profile was different: fewer Egr-1 KO B cells gave rise to CD138+ plasma cells than WT B cells. This effect occurred although Egr-1 KO B cells expressed Egr-2 and -3 at higher levels than WT cells. To confirm this result in vivo, we immunized WT and KO mice with NP-KLH plus alum and measured aspects of the anti-NP antibody production. Anti-NP antibody titer was significantly lower in the serum from KO mice than that from WT mice. Cell numbers secreting anti-NP antibodies were less abundant in both spleen and bone marrow from KO mice than from WT. furthermore, the blimp1 expression of Egr1 KO B cells was decreased. But pax5 expression of differentiated plasma cell in Egr1 KO mice was increased. Thus, our results demonstrate that Egr-1 is non-redundantly crucial for B cells to commit toward plasma cell fates. Keywords: Egr1, Plasma cells, B cells Keywords: Sanroque, Sjögren syndrome, Mouse model 64 The 2014 Fall Conference of the Korean Association of Immunologists Immune Cell Development, Differentiation and Function P-025 Innate Lymphoid Cells Facilitate NK Cell Development Through a Lymphotoxin-Mediated Stromal Microenvironment 1,2 1 1 Tae-Jin Kim , Vaibhav Upadhyay , Vinay Kumar , 1,2 1 Kyung-Mi Lee *, Yang-Xin Fu * 1 Department of Pathology, The University of Chicago, Chicago, IL 60637, Global Research Lab, Department of Biochemistry and Molecular Biology, Korea University College of Medicine, Seoul, Korea Tel: 02-920-6253, E-mail: [email protected] P-026 Poster Presentation The Advent of Soluble Common Gamma Chain Rocks the T Cell World: A Novel Therapeutic Target for Autoimmune Diseases Changwan Hong* Department of Anatomy, Pusan National University School of Medicine, Yangsan, Korea Tel: 051-510-8041, E-mail: [email protected] 2 Natural killer (NK) cell development relies on signals provided from the bone marrow (BM) microenvironment. It is thought that lymphotoxin (LT) α1β2 expressed by the NK cell lineage interacts with BM stromal cells to promote NK cell development. However, we now report that a + small number of RORγt innate lymphoid cells (ILCs), and not + −/− CD3 NK1.1 cells, express LT to drive NK development. Similar to LT or RORγt−/− mice, the mice conditionally lacking LTα1β2 on RORγt+ ILCs experience a developmental arrest at the immature NK stages, between stages of NK development to the mature NK cell stage. This developmental block results in a functional deficiency in the clearance of + NK-sensitive tumor cells. Reconstitution of Thy1 ILCs from BM or puri+ fied RORγt ILCs from lamina propria lymphocytes into LT-deficient RORγt+ BM cultures rescues NK cell development. These data highlight + a previously undiscovered role of RORγt ILCs for NK cell development and define LT from ILCs as an essential molecule for the stromal microenvironment supporting NK cell development. The common γ-chain (γc) plays a central role in signaling by IL-2 and other γc−dependent cytokines. Here we report that activated T cells produce an alternatively spliced form of γc that results in secretion of the γc extracellular domain. The soluble form of γc (sγc) was produced only by alternative splicing and not receptor shedding, and directly bound to IL-2Rβand IL-7Rαproteins on T cells to inhibit cytokine signaling and promote inflammation. Soluble γc impaired naïve T cell survival by suppressing IL-7 signaling during homeostasis and enhanced Th17-mediated inflammation by inhibiting IL-2 signaling upon T cell activation, as sγcoverexpressing mice are consequently more susceptible to EAE. Thus, sγc expression is a new mechanism for regulating cytokine signaling and controlling T cell activation and differentiation. Furthermore, these data may lead to the generation of novel therapeutic targets for the treatment of inflammatory autoimmune diseases, as sγc is genetically conserved in human and circulating levels of sγc have been linked to autoimmune diseases patients. Keywords: Th17, Autoimmune disease, IL-2, IL-7, Common gamma chain, T cell differentiation Keywords: NK cells, ILC3, RORγt, Lymphotoxin P-027 The Identification of Transcription Factors Involved in Function and Development of Innate CD8 T Cell by the GeneMine Byunghyuk Lee, Changwan Hong* P-028 Migration Property of Regulatory T Cells in Human Acute Viral Infection Jeewon Lee1, Yoon Seok Choi2, Eui-Cheol Shin1* 1 Department of Anatomy, Pusan National University School of Medicine, Yangsan, Korea Tel: 051-510-8041, E-mail: [email protected] Generally, many studies clearly defined the basic parameters for the matura+ + + + tion of conventional naïve CD8 and CD4 T cells from CD8 CD4 DP thymocytes, which included a dependence on TCR interactions with allele-specific MHCI or MHCII, respectively, a lack of high-affinity self-reactivity and a dependence on co-receptor engagement. Until now, the development of DP thymocytes into mature T cells was thought to involve this single, binary decision process. However, it is now apparent that DP thymocytes give rise to many lineages of mature T cells, in addition to conventional naïve CD8+ and CD4+ T cells. The two best-studied examples of these alternative, non-conventional lineages are Treg, NKT cells, and the other example is CD8+ T cells which are restricted with non-classical MHCIb, Qa-1 and MR1. Interestingly, most of studies have identified proteins that are crucial for the development of Treg or NKT cell lineages. Recent reports showed that TEC kinases are required for the development of conventional naïve CD8+ T cells, but repress the alternative subset of non-conventional CD8+ T cells such as innate CD8+ T cells. Innate CD8+ T cells have been characterized by specific transcription factor, high Eomesodermin (Eomes) and low Tbet. However, the key signaling pathways and transcription factors that promote conventional vs innate CD8+ T cell development have not been defined. And also the reasons why development of these cells should be inhibited need to be defined. To find which specific genes are involved in function and development of innate CD8+ T cells, we first investigated the frequency of innate CD8+ T cells and NKT cells in different strains of mice. In further study, specific target genes of innate CD8+ T cells will be evaluated by ‘The GeneMine’ method. The information will provide the key target genes responsible for the conserved aspects of all innate T cell subset, of which the most important is their ability to rapidly respond to activation signals without requiring extensive priming and expansion. Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, KAIST, 2Department of Internal Medicine, Chungnam National University College of Medicine, Daejeon, Korea Tel: 042-350-4266, E-mail: [email protected] + FoxP3 regulatory T cells (Tregs) play a major role in maintaining homeostasis of the immune system. Disruption in the development or function of Tregs causes autoimmune diseases in human and animals. To function properly, Tregs need to manipulate their migration ability into specific inflamed site and suppressive activity to control effector T cells. In this study, we focused on migration property of Tregs and investigated which chemokine receptors are expressed by Tregs during acute hepatitis A (AHA) which is caused by hepatitis A virus (HAV) infection. In the present study, we investigated chemokine receptors of Tregs using peripheral blood mononuclear cells (PBMCs) of AHA patients and compared those in acute and convalescence phase as well as healthy donors. The frequency of CCR4+ and CXCR3+ Tregs showed no difference between AHA patients and healthy donors. In the case of CCR4, its expression level on Tregs was rather decreased in AHA patients compared to healthy donors. On the contrary, the expression of CCR5+ Tregs was significantly increased in AHA patients compared to healthy donors. In addition, CCR5 ligand chemokines were increased in the sera of AHA patients, and their secretion by HAV-infected cells was observed in an in vitro infection system. Taken together, these data suggest that CCR5 might play a crucial role in the peripheral homing of Tregs to the inflamed liver. Keywords: Regulatory T cells, Acute hepatitis A, C-C chemokine receptor type 5 Keywords: Innate CD8+ T cells, NKT cells, Eomes, The GeneMine The 2014 Fall Conference of the Korean Association of Immunologists 65 Poster Presentation P-029 Immune Cell Development, Differentiation and Function Functional Heterogeneity of Influenza + Virus-specific CD8 T Cells 1 2 1 Jihye Kim , Hyeok-Il Kwon , Jeewon Lee , Min-Suk 2 2 1 Song , Young Ki Choi , Eui-Cheol Shin * 1 Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology, Daejeon, 2College of Medicine and Medical Research Institute, Chungbuk National University, Cheongju, Korea Tel: 042-350-4266, E-mail: [email protected] CD8+ T cells clear viruses through their two main effector functions: cytolysis and cytokine secretion. However, the factors controlling these antiviral effector activities in vivo at infection sites are poorly understood. In this study, we suggest that IL-15 induces functional heterogeneity of anti+ viral CD8 effector T cells in the influenza virus-infected lungs. We stud+ ied CD8 T cell responses in a mouse influenza model, using highly virulent (ma81) and its single amino acid substitution (a Iso -to- Thr at position 97) variant, avirulent (ma81-PAI97T) influenza virus strains. We found + that CD8 T cells from the lethal influenza (ma81)-infected lungs exerted strong cytotoxicity, evidenced by CD107abright, without IFN-γ secretion bright − + IFN-γ ). In contrast, conventional effector CD8 T cells (CD107a dim + with IFN-γ secretion and moderate cytotoxicity (CD107a IFN-γ ) were observed in the avirulent influenza-infected lungs. Interestingly, the amount of IL-15 was increased in the BAL fluid during the lethal infection, and in vivo anti-IL-15 blocking reduced the frequency of CD107abrightIFN-γ− subset, suggesting that CD107abrightIFN-γ− CD8+ T cells are generated by IL-15. These results indicate the importance of + IL-15 on functional heterogeneity of CD8 T cells to mount appropriate responses in the context of influenza infection. P-030 CD244-CD48 Interaction on Murine iNKT Cells is Critical for the Optimal Activation of iNKT Cells Jung Hoon Shin, Se-Ho Park* Department of Life Sciences, Korea University Tel: 02-3290-3160, E-mail: [email protected] iNKT cells are a specialized subset of T cells that modulate immune responses in diverse ways and that require fine regulation of their own reactivity. CD244 (2B4) is a member of SLAM family of molecules and can act as both an inhibitory and an activating receptor, depending on the relative expression levels of 2B4 and its ligand, CD48. Although iNKT cells are known to express 2B4, the function of 2B4 on iNKT cells remains largely undefined. Here, we report that 2B4 is required for optimal activation of iNKT cells. In 2B4 knock-out (KO) mice, iNKT cells showed impaired activity, and the symptoms of concanavalin A (Con A)-induced hepatitis were substantially weaker in 2B4 KO mice than in wild-type mice. We demonstrate that CD48 engagement is also required for the function of 2B4 in iNKT cells. Strikingly, however, CD48 providers were not necessarily target cells, but rather the iNKT cells themselves. CD48 expressed on iNKT cells directly engaged 2B4 on iNKT cells through homotypic cell-to-cell interactions. Our data highlight the central role of 2B4/CD48 in homotypic iNKT cell-to-cell interactions and demonstrate the requirement of 2B4/CD48 for optimal activation both in vitro and in vivo models of acute inflammation. Keywords: 2B4, CD48, iNKT cells + Keywords: Influenza virus, CD8 T cells, Cytotoxicity, IFN-γ, IL-15 P-031 Gut Microbiota Controls Homeostasis of Hematopoietic Stem Cells through Toll-like Receptor Signaling 1 1 1 Seungwon Lee , Ohseop Kwon , Hyekang Kim , Young 1 3 1 Min Kim , Kwang Soon Kim , Yunji Park , You-Me 1,2 3 1,2 Kim , Charles D. Surh , Seung-Woo Lee * 1 Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology, 2Department of Life Sciences, Pohang 3 University of Science and Technology, Academy of Immunology and Microbiology, Institute for Basic Science, Pohang, Korea Tel: 054-279-2355, E-mail: [email protected] Homeostasis of hematopoietic stem cells (HSCs) is important for maintaining their activity in response to physiological stresses or demands throughout one’s lifespan. However, it is not well characterized which signals are involved in regulating homeostasis of HSCs. Here, we revealed for the first time that gut microbiota is capable of controlling the homeostasis of HSCs. Conventional SPF mice − which were treated with antibiotics showed decreased total number of Lineage SCA-1+c-KIT+ cells (LSKs), including HSCs and multipotent progenitor cells (MPPs), in the bone marrow (BM). In particular, the number of short-term HSCs, which represent more activated form of HSCs, was significantly decreased, while that of long-term HSCs was not changed. Cell cycle analyses have shown that HSC turnover was markedly reduced in antibiotics treated mice. In parallel, germ-free (GF) mice showed the altered homeostasis of HSCs with attenuated cell cycle progression, which was restored by re-colonization of gut microbiota. These results indicate that gut microbes indeed play a role in controlling homeostasis of HSCs through the regulation of cell cycle progression. Since commensal microorganisms are recognized by pattern recognition receptors, such as −/− Toll-like receptors (TLRs), we analyzed HSC homeostasis in MyD88 and 3d mice, both of which are defective in many TLR signaling pathways. Consistent −/− with previous results, both MyD88 and 3d mice showed decreased number of HSCs, especially short-term HSCs, implying that TLR signaling contributes to homeostasis of HSCs. HSC analyses with mice lacking of specific TLR suggest that TLR5 and TLR9, but not TLR3, are involved in HSCs homeostasis. In sum, our results reveal the as-yet-unknown link between colonization of commensal microbes at the gut and homeostasis of HSCs at the BM, providing a new perspective on the evolution of host-commensal microorganism interactions. Keywords: Gut microbiota, Hematopoietic stem cell (HSC), Toll-like receptor (TLR) 66 P-032 Effect of Estrogen Treatment on Macrophage Populations in Mouse Adipose Tissue Kyeongdae Kim1, Jeong Min Choe2, Junhee Choi1, Young Jin Wi1, Jae-Hoon Choi1*, Hyun Tae Park2* 1 Laboratory of Pathophysiology, Department of Life Science, Hanyang University, 2Department of Obstetrics and Gynecology, College of Medicine, Korea University, Seoul, Korea Tel: 02-2220-2540, E-mail: [email protected] During the progression of obesity, large number of immune cells has been shown to be accumulated in adipose tissue including dendritic cell (DC), macrophages, T cells, B cells, and regulatory T cells (Tregs). Especially, obesity increases the number of CD11c+ F4/80+ macrophage, which is M1 or classically 1 activated macrophage. In human, inactivating mutations in the estrogen receptor alpha isotype cause a metabolic syndrome and obesity in woman.2,3 E2 has also been known to affect appetite, and adipose tissue macrophages (ATMs). To understand the effect of E2 on ATMs in high fat diet-fed mice, we treated E2 to ovariectomized mice using osmotic pump, and compared the E2 effect on ATMs from free-feeding and pair-feeding mice. E2 treatment significantly increased M2 macrophages in adipose tissue, while M1 macrophage was not affected by E2. To understand the mechanism responsible for the increased M2 ATMs by E2 treatment, we first analyzed the Ki67 expression on the M2 ATMs, and found that the Ki67 level of M2 ATMs was not changed by E2 treatment. Furthermore, EdU was injected to the mice having depleted blood monocytes by clodronate injection. EdU+ M2 percentage on E2 group is not significantly increased compared to vehicle group. Those results indicate that E2 increases the M2 ATMs not by local proliferation, but by inducing more recruitment of blood monocytes into the tissue. References 1. Morris DL, Cho KW, Delproposto JL, et al. Diabetes 2013;62:2762-2772 2. Meyer MR, Haas E, Barton M. Gender differences of cardiovascular disease: new perspectives for estrogen receptor signaling. Hypertension 2006;47:1019-1026 3. Steinbaum SR. The metabolic syndrome: an emerging health epidemic in women. Progress in Cardiovascular Disease 2004;46:321-336 Keywords: Obesity, Adipose tissue, Macrophage, Estrogen The 2014 Fall Conference of the Korean Association of Immunologists Immune Cell Development, Differentiation and Function P-033 The Effect of FoxO6 on Dendritic Cell P-034 Mi Eun Kim, Jun Sik Lee* Poster Presentation IL-33 Induces Th17 Cell Differentiation through Increased IL-6 Production in Bone Marrow-derived Dendritic Cells Department of Biology, Immunology Research Lab, BK21-plus Research Team for Bioactive Control Technology, College of Natural Sciences, Chosun University, Gwangju, Korea Tel: 062-230-7459, E-mail: [email protected] Su-Ho Park, Tae Sung Kim* Forkhead box O (FoxO) is transcription factors which is involved in cell cycle progression, cell survival and DNA-repair. Dendritic cells (DC) are professional antigen-presenting cells which present the antigens to T lymphocytes and regulate the adaptive immunity. Previous studies indicated that FoxO 1 and 3 are plays a critical role in DC maturation and DC-T cell interaction. FoxO6, a member of FoxO family, is less attended than other FoxO families, because limited expression region of FoxO6. Also, the effect of FoxO6 is still unknown. Here, we investigated the effect of FoxO6 in dendritic cells. Out data suggest that FoxO6 is expressed in murine dendritic cells. Moreover, phosphorylated FoxO6 is increased and acetylated FoxO6 is decreased during dendritic cell differentiation. We expect that FoxO6 has important role in DC differentiation and maturation, and will further investigate the functions and the mechanisms of FoxO6 on DC. Interleukin (IL)-33, a member of the IL-1 cytokine family and a ligand for the complex of membrane-bound ST2L and IL-1R accessory protein, has been associated with a variety of autoimmune diseases, such as sclerosis, inflammatory bowel disease, and rheumatoid arthritis. Autoimmune diseases are related to activation of Th17 cells and deficiency of Treg cells. The IL-33-activated bone marrow-derived dendritic cells (BMDCs) are known to involve in the differentiation of Th2-type cytokines in CD4+ T cells. However, the effects of IL-33 on Th17 cells via BMDCs remain clearly unknown. In this study, we investigated the effects of IL-33 on BMDC-mediated CD4+ T cell polarization and the underlying signaling pathways. Treatment of BMDCs with IL-33 up-regulated expression of b cell-surface molecules CD40, CD80, and I-A (MHC class II), markers of DC maturation. IL-33 increased production of IL-6, which is known as one of the Th17-polarizing cytokines, in BMDCs. Importantly the + IL-33-treated DCs enhanced the differentiation of CD4 T cells into IL-17-producing Th17 cells, while they reduced the differentiation into IL-10-producing Treg cells. Interestingly, both array data and RT-PCR show that IL-33 induces the expression of suppressor of cytokine signaling-3 (SOCS-3) in BMDCs. The research of signaling pathway about Th17 cell differentiation depending on IL-33-stimulated BMDCs is under investigation. The results of this study will be helpful to understand immunopathologic roles of IL-33 in severe autoimmune diseases. Keywords: FoxO6, Dendritic cells, DC maturation, DC differentiation Department of Life Sciences, Korea University, Seoul, Korea Tel: 02-3290-3920, E-mail: [email protected] Keywords: Interleukin-33, Bone marrow-derived dendritic cell, Th17 cell, SOCS-3, Autoimmune disease P-035 PlGF-1 and -2 Induce Hyperplasia and Invasiveness of Primary Rheumatoid Synoviocytes Seung-Ah Yoo1, Ji-Hwan Park2, Seoung-Hye Hwang1, 1 1 3 Sang-min Oh , Sasung Lee , Valeria Cicatiello , Sangchul 4,5 3,6 4,5 Rho , Sandro De Falco , Daehee Hwang , Chul-Soo Cho1,7, Wan-Uk Kim1,7* 1 POSTECH-CATHOLIC BioMedical Engineering Institute, The Catholic University of Korea, Seoul, 2Department of Chemical Engineering, POSTECH, Pohang, Korea, 3Istituto di Genetica e Biofisica “Adriano Buzzati-Traverso”, National Research Council, 80131 Napoli, Italy, 4 School of interdisciplinary bioscience and bioengineering, POSTECH, Pohang, 5Department of New Biology and Center for Plant Aging Re6 search, Institute for Basic Science, DGIST, Daegu, Korea, Bio-Ker, MultiMedica Group, 80131, Naples, Italy, 7Department of Internal Medicine, The Catholic University of Korea, Seoul, Korea Tel: 02-2258-7831, E-mail: [email protected] Inflammation-mediated oncogenesis has been implicated in a variety of cancer types. Rheumatoid synovial tissues can be viewed as a tumor-like mass, consisting of hyperplastic fibroblast-like synoviocytes (FLSs). FLSs of rheumatoid arthritis (RA) patients have pro-migratory and invasive characteristics, which may be caused by chronic exposure to genotoxic stimuli, including hypoxia and growth factors. Here, we tested whether a transformed phenotype of RA-FLSs are associated with placental growth factor (PlGF), a representative angiogenic growth factor induced by hypoxia. We identified that PlGF-1 and PlGF-2 are the major PlGF isoforms in RA-FLSs. Global gene expression profiling revealed that cell proliferation, apoptosis, angiogenesis, and cell migration were mainly represented by differentially expressed genes in RA-FLSs transfected with siRNA for PlGF. Indeed, PlGF-deficient RA-FLSs showed a decrease in cell proliferation, migration, and invasion, but an increase in apoptotic death in vitro. To the contrary, PlGF gene overexpression resulted in the opposite. Moreover, exogeneous PlGF-1 and PlGF-2 increased survival, migration, and invasiveness of RA-FLSs by binding their receptors, Flt-1 and NP-1, up-regulating the expression of anti-apoptotic molecules, including pErk and Bcl2. In a xeno-transplantation model, knock-down of PlGF transcripts reduced RA-FLS proliferation and migration. Collectively, in addition to their role for neovascularization, PlGF-1 and -2 promote proliferation, survival, migration, and invasion of RA-FLSs in an autocrine and paracrine manner. These results provide evidences how primary cells with mesenchymal origin acquire aggressive and transformed phenotype, suggesting that PlGF and its receptors offer new targets for an ant-FLS therapy. P-036 The Expression of CD138 Is Regulated Via Transcription and Shedding of Transmembrane Protein in B Cells Joo Hee Lee, Chanho Park, In Seong Kho, Tae Jin Kim* Department of Molecular Cell Biology and Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, Suwon, Korea Tel: 031-299-6161, E-mail: [email protected] CD138 is known as a marker of plasma cells in B cell lineage, but an intermediate level of the CD138 expression was recently reported in the pre-B cells and a subset of follicular B cells. We previously identified a CD138+IgMlowIgDhigh B cell population that belonged to follicular B-2 cells. Here we investigated the regulatory mechanism of the CD138 expression and found that the expression of CD138 was regulated at the transcriptional level as well as via a rapid shedding of transmembrane proteins. Firstly, the CD138+ B cells showed a higher mRNA expression of CD138 than the CD138- B cells, suggesting that the expression of CD138 was regulated transcriptionally. The CD138 mRNA expression was upregulated by the stimulation with PMA. Secondly, the CD138 expression was found in the presumably autoreactive B cells. We observed that CD138+ B cells belonged to follicular type I B cells or T3 transitional B cells, which suggest that CD138+ B cells are autoreactive. Interestingly, B cells from anti-hen egg lysozyme (HEL) immunoglobulin gene transgenic mice did not express CD138, but the expression of CD138 was upregulated by the in vivo injection of HEL. Notably, the CD138 expression was rapidly downregulated by the shedding as the CD138 downregulation was blocked by the pretreatment with protease inhibitors. The stimulation of B cells with 5 ng/ml PMA was sufficient for the shedding of CD138. In summary, we suggest that the expression of CD138 is regulated in a complex manner in B cells and that CD138+ B cells are a newly identified autoreactive B cell population. It is needed to investigate how a high expression of CD138 is achieved in the course of plasma cell differentiation. Keywords: CD138, Follicular B cell, PMA, HEL-tg mice Keywords: Fibroblast-like synoviocyes, Rheumatoid arthritis, Placental growth factor The 2014 Fall Conference of the Korean Association of Immunologists 67 Poster Presentation P-037 Immune Cell Development, Differentiation and Function Mst1 Deficiency Increases Natural Killer Cell population by Enhancing the Transition of pNK to iNK in the Lymphoid Organs Kyung-Min Cho, Tae Sung Kim* Department of Life Sciences, College of Life Sciences and Biotechnology, Korea University, Seoul, Korea Tel: 02-3290-3920, E-mail: [email protected] Natural killer (NK) cells are the one of the lymphocyte subset distinct from T and B lymphocytes, playing a critical role in viral immunology and anti-tumor immunotherapy. Many regulatory factors are known to be involved in NK cell development and maturation. Mst1, mammalian STE20-like kinase1, is a multifunctional serine/threonine kinase and highly expressed in immune organs, such as spleen and lymph node in which sequentially developmental processes of NK cells take place. Mst1 plays important roles in cell proliferation, differentiation, apoptosis and organ size regulation. However, it is not clear whether Mst1 plays a role in NK cell differentiation and development. Here, we investigated the role −/− of Mst1 in the development of NK cells using conventional Mst1 mouse. In contrast to the reduced numbers of T cells in Mst1−/− mouse, the frequency and numbers of NK cell population were significantly in−/− creased in the secondary lymphoid organs of Mst1 mouse. Especially, immature NK cell (iNK) and c-kit+ NK cell populations were increased in −/− Mst1 mouse, compared to those in littermate wildtype mouse. We also −/− reveal that the increased numbers of iNK cell population in Mst1 mouse were not attributed to defect in the differentiation into mature NK cells. Therefore, Mst1-deficiency enhanced the generation of iNK cells −/− resulting in the increased numbers of NK cells in Mst1 mouse. Collectively, these findings suggest that Mst1 negatively regulates the differentiation of mouse NK cells at the transition of pNK to iNK. P-038 Regulation of Mucosal and Systemic IgA Response by γδ T Cells 1 1 1 Hyeon-Jin Kim , Hye-Ju Han , Seong-Ho Kang , Bo-Ra 1 2 2 3 Jin , Aeri Shim , Hyun-Jeong Ko , Sung-Gyoo Park , 4 1 Woan-Sub Kim , Pyeung-Hyeun Kim * 1 Department of Molecular Bioscience, College of Biomedical Science, Kangwon National University, 2Laboratory of Microbiology and Immunology, College of Pharmacy, Kangwon National University, Chuncheon, Korea, 3School of Life Sciences, Gwangju Institute of Science and Technology (GIST), Gwangju, 4Division of Animal Life and Environmental Science, Hankyong National University, Korea Tel: 033-250-8546, E-mail: [email protected] γδT cells play an important role in gut mucosal immunity. Several studies showed that mucosal IgA response is impaired in γδT cell-deficient mice. Nevertheless, the role of γδT cells in the synthesis of Ig isotype re−/− mains unclear. We compared Ig expression in TCRδ mice with WT mice. Opposing to the previous report, total IgA production and surface IgA expression are elevated in TCRδ−/− mice. Likewise, germ-line α (GLα) transcript, an indicative of IgA class switching recombination −/− (CSR), was higher in PP and MLN from TCRδ mice. We also found that IgA production by peritoneal B1 cells from TCRδ−/− mice was greater than from WT mice. In addition, this increase of IgA was further augmented by treatment of retinoic acid, lactoferrin, or TGF-β1. Finally, the frequency of eosinophils in lamina propria of TCRδ−/− mice was higher than that of WT mice, supporting the early report that eosinophil maintains IgA plasma cells. Collectively, our data reveal that γδT cells affect the early phase of B cells to commit IgA isotype switching in mice. Keywords: γδT cell, IgA, B1 cell, Eosinophil Keywords: Mst1, NK cells, Mouse, Development, Differentiation P-039 PLA2 Inhibits on Acetaminophen-Induced Acute Liver Injury by Modulating Regulatory T Cells and IL-10 in Mice Hyunseong Kim1, Hwan-Suck Chung1, Hyunsu Bae1,2* 1 Department of Physiology, College of Korean Medicine, Kyung Hee University, 2Institute of Korean Medicine, Kyung Hee University, Seoul, Korea Tel: 02-961-0323, E-mail: [email protected] The aim of this study was to investigate the protective effects of phospholipase A2 (PLA2) from bee venom against acetaminophen-induced hepatotoxicity through CD4+CD25+Foxp3+ T cells (Treg) in mice. Acetaminophen (APAP) is a widely used antipyretic and analgesic, but an acute or cumulative overdose of acetaminophen can cause severe hepatic failure. Tregs have been reported to possess protective effects in various liver diseases and kidney toxicity. We previously found that bee venom strongly increased the Treg population in splenocytes and subsequently suppressed immune disorders. More recently, we found that the effective component of bee venom is PLA2. Thus, we hypothesized that PLA2 could protect against liver injury induced by acetaminophen. To evaluate the hepatoprotective effects of PLA2, C57BL/6 mice or interleukin-10-deficient (IL-10−/−) mice were injected with PLA2 once a day for five days and sacrificed 24 h after acetaminophen injection. The blood sera were collected 0, 6, and 24 h after acetaminophen injection for the analysis of aspartate aminotransferase (AST) and alanine aminotransferase (ALT). PLA2-injected mice showed reduced levels of serum AST, ALT, proinflammatory cytokines, and nitric oxide (NO) compared with the PBS-injected control mice. However, IL-10 was significantly increased in the PLA2-injected mice. These hepatic protective effects were abolished in Treg-depleted mice by antibody treatment and in IL-10−/− mice. Based on these findings, it can be concluded that the protective effects of PLA2 against acetaminophen-induced hepatotoxicity can be mediated by modulating the Treg and IL-10 production. Keywords: Phospholipase A2, Regulatory T cells, Acetaminophen, Hepatotoxicity 68 P-040 Ca2+/Hypoxia Enhances Proliferation and Immunosuppression Capacity in Human Umbilical Cord Blood-Derived Marrow Stromal Cells Yu Jin Hong, Mi-Yeon Lee, Hong Bae Jeon, Su Jin Choi, Wonil Oh, Yoon-Sun Yang, Soon-Jae Kwon, Eun Su Jeon* Biomedical Research Institute, MEDIPOST Co., Ltd., Seoul, Korea Tel: 82-2-3465-6771, E-mail: [email protected] Umbilical cord blood-derived marrow stromal cells (UCB-MSCs) with high proliferation capacity and immunomodulatory properties are considered to be a good candidate for cell-based therapies. But until now, little work has been focused on the immunosuppression capacity of hUCBMSCs. Inflammatory cytokines are critical in inducing the immune modulatory properties of MSCs. However, the detailed regulatory mecha+ nisms are yet to be fully understood. Ca2 /Hypoxia-conditioned hUCB-MSCs (CMH-MSCs) were evaluated that the immunogenicity and immunosuppressive activity was lower than naïve hUCB-MSCs. The functionality of the engrafted human immune cells was verified using a mixed lymphocyte reaction (MLR) to measure T cell proliferation. Furthermore, we showed that the effect of CMH-MSCs is exerted through promoting anti-inflammatory cytokines induced PGE2 expression. Additionally, CMH-MSCs showed higher effects on the proliferation of MSCs than using calcium and hypoxia individually. Emphysema is characterized by alveolar destruction and persistent inflammation of the airways. In a model of elastase-induced emphysema we examined the therapeutic effect of CMH-MSCs. Adult mice developed emphysematous changes in the lung tissue on day 21 after elastase treatment, whereas emphysematous changes were decreased in CMH-MSCs-injected mice compared to naïve-injected mice. Altogether, these data suggest that treat+ ment with Ca2 /Hypoxia in hUCB-MSCs enhanced the immunosuppressive capacity of hUCB-MSCs. High capacity hUCB-MSCs a very useful model for clinical application of allogeneic cell therapies. Keywords: UCB-MSCs, Ca2+/Hypoxia, Emphysema The 2014 Fall Conference of the Korean Association of Immunologists Immune Cell Development, Differentiation and Function P-041 Chitinase Like Protein BRP-39 Inhibits Th1 Differentiation Through Regulation of IFNγ Mediated STAT1 Signaling Pathway P-042 Poster Presentation Protein Kinase CK2 Mediates SIN-1-Induced Heme Oxygenase-1 Expression in Articular Chondrocytes Do-Hyun Kim1,2, HongJai Park1,2, JaeUng Lee1, Je-Min Choi1,2* Da Sol Kim1,2, Kang Mi Kim2, Mi Sun Lee2, Young Chul Park2* 1 Department of Life Science, 2Research Institute for Natural Sciences, Hanyang University, Seoul, Korea Tel: 02-2220-4765, E-mail: [email protected] 1 Department of Physiology, 2Department of Microbiology & Immunology, Pusan National University School of Medicine, Yangsan, Korea Tel: 051-510-8093, E-mail: [email protected] Chitinase is evolutionary conserved enzyme that involved to regulation of parasite life cycle and anti-parasitic immune response. Chitinase like protein (CLP), which lack a chitinase activity by mutations of enzyme site also participated to anti-parasitic innate immunity, but their roles in T cell immunity has not been studied. Here, we investigated the role of mouse −/− mouse CLP BRP-39 in T cell immune response by using BRP-39 model. BRP-39 mRNA is expressed in lymphoid tissue and T cells with its up-regulation upon TcR or IL-4 stimuli. BRP-39 protein is secreted from lineage specific T cell subsets, especially Th17 cells. Although BRP-39−/− mouse does not have significant phenotype, BRP-39−/− T cells showed increased production of IL-2 and IFN-γ with hyper-pro−/− naïve T cells sigliferative response TcR stimuli. In addition, BRP-39 nificantly more strongly differentiated into Th1 cells with increase of T-Bet and Runx3 while their differentiation into Th2 was reduced with lower IL-4, 5 production and JunB expression. Also, pSTAT1, 4 were increased in BRP-39−/− Th1 cells while their Th2 cells showed lower pSTAT6 than WT. We found that expression of PTPN2, a phosphatase −/− which regulate STAT1 phosphorylation was reduced in BRP-39 Th1 cells and recombinant BRP-39 treatment could not reverse the phenotype suggesting intrinsic BRP-39 down-regulate IFNγ expression via regulation of IFNγ/STAT1 signaling pathway. Collectively, our results demonstrate for the first time novel regulatory roles of BRP-39 in T cell activation and differentiation, and suggest possibility of novel therapeutic target for regulation of Th1 related autoimmune diseases. Heme oxygenase-1 (HO-1) is induced as an adaptive mechanism against oxidative stress in chondrocytes, which play an important role in the maintenance and degradation of cartilage. In the present study, we examined the role of protein kinase casein kinase (CK2) on peroxynitrite-induced expression of HO-1 in primary articular chondrocytes. 3-Μorpholinosydnonimine hydrochloride (SIN-1) has been shown to mediate cell death by activating apoptosis-related molecules in cells. In this study, we used a low concentration of SIN-1 that did not induce apoptosis to elucidate the mechanism by which SIN-1 upregulates HO-1 expression. In chondrocytes, SIN-1 induced HO-1 expression with spontaneous downregulation in a different manner than with high concentrations of SIN-1. Importantly, SIN-1 treatment of chondrocytes increased CK2 activation. Additionally, inhibition of CK2 with 4,5,6,7-tetrabromobenzotriazole (TBB) or siRNA did not induce HO-1 expression and reduced NF-E2-related factor 2 (Nrf2) accumulation in chondrocytes. Therefore, we examined whether CK2 directly regulates Nrf2, which is a transcription factor that regulates the expression of HO-1. Indeed, TBB treatment inhibited phosphorylation and nuclear translocation of Nrf2 in SIN-1-treated cells. Moreover, using an immunoprecipitation assay, we confirmed that SIN-1 treatment enhanced the interaction between CK2 and Nrf2. Taken together, our findings suggest that peroxynitrite activates Nrf2 via CK2 signaling, leading to the upregulation of HO-1 in primary chondrocytes. Keywords: Chitinase/CLP, Th1/Th2 balance, STAT signaling pathway, PTPN2, Autoimmune disease Keywords: HO-1 expression, Peroxynitrite, Protein kinase CK2, Nrf2, Chondrocytes P-043 Identification of Liver Homing Memory T Cells in Response to Acute Viral Infections P-044 Hyun Gyung Kim, Yong Woo Jung* Department of Pharmacy, Korea University, Sejong, Korea Tel: 041-860-1618, E-mail: [email protected] Acute viral infections activate pathogen-specific CD8+ T cells that give rise to long-lived memory cells. These memory T cells exist in the lymphoid organs as well as the non-lymphoid organs. Of particular interest is the liver because it is home for numerous pathogens. In addition, the liver contains surprisingly large number of memory T cells. Therefore, we hypothesized that the liver provides important factors for the homeostasis of memory T cells. In order to test this hypothesis, we infected mice with lymphocytic choriomeningitis virus and dissected the liver on day 15 post infection. Using 4-color immunofluorescent microscopy, virus-specific memory precursor effector cells were found in the “lymphoid aggregates”. In order to examine if these cells are actively homing to the liver, we isolated memory T cells from the liver and spleen, and transferred them to another mouse intravenously. These cells can migrate into both the spleen and liver, but they preferentially home to the liver compared to the cells obtained from the spleen. The liver-homing memory T cells showed the low expression of CD27 and CD122. Taken together, memory CD8 T cells were found in the lymphoid aggregates of the liver, and they were preferentially recruited into this site. Keywords: Acute viral infection, Lymphocytic choriomeningitis virus, Memory precursor effector cells, Lymphoid aggregates IL-7Rαlow Memory CD8+ T Cells Are Clonal Anergic Cells in Human and Increased in Patients with Common Variable Immunodeficiency 1† 1,2† 1† Ji Hyun Sim , Jin-Hee Kim , Bon-A Cho , Youngho 3 1 1,2 1,2 Ko , Sun-Kyung Lee , Min A Seol , Keunhee Oh , Dong-Sup Lee1,2, Joong Gon Kim5, Ae Kyung Park6, Insoo 7 2,4 1,2 Kang , Kyung-Ho Choi , Hang-Rae Kim * 1 Department of Anatomy, 2Department of Biomedical Sciences, 3Depart4 ment of Microbiology and Immunology, and Department of Biochemistry and Molecular Biology, 5Department of Pediatrics, Seoul National University College of Medicine, Seoul, 6College of Pharmacy, Sunchon 7 National University, Suncheon, Korea, Section of Rheumatology, Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520, USA † The first three authors equally contributed to this work. Tel: 02-740-8214, E-mail: [email protected] We have found two subsets of CD8+ T cells expressing IL-7Rαhigh and IL-7Rαlow with different cell survival responses to IL-7 in peripheral blood of human. Of interest, these IL-7R αlow memory CD8+ T cells that comparably express effector molecules have impaired proliferation and IL-2 production upon TCR stimulation by anti-CD3/CD28 Abs. But the mechanism for this is unknown. Treating IL-7Rαlow memory CD8+ T cells with anti-CD3/CD28 Abs did not produce IL-2, but comparably expressed IFN-γand TNF-α, as shown in split anergic cells. Importantly, IL-7Rαlow memory CD8+ T cells have reduced calcium flux and subsequent inhibited nuclear translocation of NF-AT1 upon in vitro TCR stimulation by anti-CD3/CD28 Abs, leading to impaired IL-2 production. Moreover, inhibition of IL-2 production and proliferation was not recovered by PMA/ionomycin treatment, suggesting that IL-7Rαlow memory CD8+ T cells also have TCR-distal signaling defect of TCR signaling. Interestingly, this anergic status was reversed by exogenous IL-2 stimulation for 10 days changing fresh media every 3 days in vitro, clearly distinct from adaptive tolerance induced by soluble peptide Ag. Notably, IL-7Rαlow memory CD8+ T cells are increased in patient with common variable immunodeficiency (CVI) exhibit an abnormal T-cell population. In conclusion, we firstly showed that specific subset of human CD8+ T cells, IL-7Rαlow memory CD8+ T cells, are T-cell clonal anergy in vivo. Also, these cells are enriched in CVI patient. Based on these findings, we are going to further investigate which mechanisms are involved in anergic phenotype of human anergic IL-7Rαlow memory + CD8 T cells, and eventually, what is the role of these cells in vitro using by MLR system. Keywords: IL-7 receptor alpha (α), Memory CD8+ T cells, Clonal anergy, CVID The 2014 Fall Conference of the Korean Association of Immunologists 69 Poster Presentation P-045 Immune Cell Development, Differentiation and Function Identification of Specialized-Microenvironments for Interleukin-7 Signaling in Various Organs Kyong Hoon Kim1, Un-Hwan Ha1, Yong Woo Jung2* P-046 Induction of Heme Oxygenase 1 Expression Protects Articular Chondrocytes Against Cilostazol Induced Cellular Senescence 1 Mi Sun Lee1, Kang Mi Kim1, Da Sol Kim1,2, Young Chul Park1* 2 1 Department of Biotechnology and Bioinformatics, Korea University, Department of Pharmacy, Korea University, Sejong, Korea Tel: 041-860-1618, E-mail: [email protected] Memory cells induce rapid immune response against secondary infection and these cells require specific cytokine-induced signals for their survival and homeostatic proliferation. It has been shown that IL-7, an important cytokine for the longevity of these cells, was expressed in several organs such as the lung, liver, lymph nodes and spleen. In order for memory T cells to see IL-7, either a few organs may secrete this cytokine systemically or specialized anatomic microenvironments also called niches may provide it locally. To test this hypothesis, we employed lymphocytic choriomeningitis virus (LCMV) to infect mice, and then analyzed the number hi of IL-7 receptor (IL-7R) cells in individual organs. Although LCMV infection induced clonal expansion of virus-specific CD8 T cells, the total numbers of IL-7Rhi cells were constant in the bone marrow, lymph nodes hi and spleen. In contrast, the number of IL-7R cells in the lung or liver was reduced following LCMV infection. We also examined the expression of IL-7 mRNA in these individual organs. Overall, the microenvironments hi may have a limit to contain certain numbers of IL-7R cells, suggesting that these organs provide IL-7 locally. Keywords: Memory cell, Interleukin-7, lnterleukin-7 receptor, Lymphocytic choriomeningitis virus Department of Microbiology & Immunology, 2Department of Physiology, Pusan National University School of Medicine, Yangsan, Korea Tel: 051-510-8093, E-mail: [email protected] Chondrocyte senescence is associated with the aging and degeneration of cartilage, and eventually leads to joint destruction. The aim of this study was to elucidate the mechanisms responsible for the cytoprotective effects of heme oxygenase-1 (HO-1) on chondrocytes in cartilage. Chondrocyte senescence was induced using cilostazol and measured using a specific senescence associated β-galactosidase (SA-β-gal) staining assay. Cilostazol altered the expression of type II collagen and β-catenin, which are phenotypic markers of the differentiation and dedifferentiation of chondrocytes. Cilostazol also significantly induced HO-1 expression, and the induction of HO-1 expression was affected by a significant increase in reactive oxygen species (ROS) production caused by cilostazol treatment. Of note, pre treatment with 3-morpholinosydnonimine hydrochloride (SIN-1), an inducer of HO-1 expression, markedly attenuated cilostazol induced chondrocyte senescence, and thus, we examined whether HO-1 directly modulates chondrocyte senescence induced by cilostazol. The upregulation of HO-1 was found to suppress cilostazol-induced cellular senescence. In addition, the inhibition of HO-1 activity with the iron chelator, desferrioxamine (DFO), or HO-1 siRNA increased cilostazol-induced chondrocyte senescence. Based on these results, it can be concluded that HO-1 is associated with the suppression of chondrocyte senescence, and that the enforced overexpression of HO-1 protects chondrocytes against stress induced senescence. Keywords: Primary articular chondrocytes, HO-1, Cilostazol, Cellular senescence, ROS P-047 Down-Regulation of Mucosal B-Cell Activating Factor by Ribosomal Inactivation P-048 Characteristics of Hypo-Responsive Subset of Naturally Occurring Tregs Kee Hun Do1, Dongwook Kim2, Yuseok Moon1* Jun Young Lee1,2, Joo Hee Kim1,2, Charles D. Surh1-3* 1 Laboratory of Mucosal Exposome and Biomodulation, Department of Microbiology and Immunology, Immunoregulatory Therapeutics Group in Brain Busan 21 Project, Pusan National University School of Medicine, Yangsan, Korea, 2National Institute of Animal Science, RDA, Suwon, Korea Tel: 051-510-8094, E-mail: [email protected] 1 Academy of Immunology and Microbiology (AIM), Institute for Basic Science (IBS), 2Department of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology (POSTECH), Pohang, Korea, 3Division of Developmental Immunology, La Jolla Institute for Allergy and Immunology, La Jolla, CA, USA Tel: 054-279-8721, E-mail: [email protected] Although the activation of B cells in the gastrointestinal tract is of great importance in the context of immunity to pathogens and mucosal inflammatory diseases, little is known about the mechanisms responsible for the local activation of B cells in the subepithelial area of the intestine. Epithelium-derived BAFF is the major modulator of B cell development and Ig class switching. The present study was performed to address the molecular mechanism of BAFF expression in gut epithelial cells in the presence of pro-inflammatory stimuli. Inflammation-induced BAFF expression in mucosal epithelial cells might be responsible for diverse mucosa-associated diseases linked to intestinal inflammation and autoimmunity. Although BAFF was marginally expressed in unstimulated epithelial cells, BAFF mRNA was significantly upregulated by pro-inflammatory IFN-γ. Furthermore, IFN-γ triggered JAK/ STAT1 signals via the cytokine receptor, which contributed to epithelial BAFF up-regulation. In terms of signaling intervention, ribosomal inactivation attenuated IFN-γ-activated JAK/STAT signal transduction and subsequent BAFF induction in gut epithelial cells. ribosomal inactivation led to the super-induction of SOCS3 by enhancing its mRNA stability via HuR RNA-binding protein. Upregulated SOCS3 then contributed to the blocking of the JAK/ STAT-linked signal, which mediated BAFF suppression by ribosomal inactivation. All of these findings show that ribosomal inactivation-induced SOCS3 plays a novel regulatory role in epithelial BAFF production, suggesting that epithelial ribosomal dysfunction in association with SOCS3 may be a promising therapeutic point in BAFF-associated human mucosal diseases This work was carried out with the support of “Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ008405032014)” Rural Development Administration, Republic of Korea. Current prevailing idea for the mechanism of Treg selection in thymus is largely relying on high TCR affinity of Tregs against self-antigen. However, investigation on the relative contribution of strong self-affinity of Tregs on maintaining Treg homeostasis in secondary lymphoid organ (SLO) remains elusive. We found that Ly6c is only expressed by naïve naturally occurring Tregs (nTreg). Interestingly, Ly6c expression on Treg is detectable in SLO but not in thymus. Furthermore, Ly6c+ Tregs were not proliferating and they exhibit impaired in vitro suppressive activity. Because expression level of indicators that associated with TCR signaling strength was not very different between Ly6c+ and Ly6c- Treg, this self-antigen hypo-responsiveness of Ly6c+ Tregs seems like associated with post-thymic developmental education against self-antigen reactivity. By analyzing Treg subsets in differentially aged mice, we found that the proportions of Treg subsets were dynamic depending on the age of mice. Ly6c+ Treg proportion is lowest at 1wk and highest at 6~8wk but the ratio is declining with aging of mice and effector Treg ratio is inversely correlates with Ly6c+ Treg ratio. To demonstrate that Ly6c Treg is thymic origin, we performed adult thymectomy and Ly6c+ Foxp3+ cells in SLO was completely depleted. Since in vitro stimulation of Ly6c+ Treg induced down-regulation of Ly6c and proliferation, the smaller ratio of Ly6c Treg in aged, young and thymectomized mice indicates that the disappearance of Ly6c Treg is associated with activation-induced down-regulation of Ly6c. Previously, we found that the rate of proliferation observed in SLO of irradiated SPF mice reflect the reactivity against self-antigen as this rate of proliferation was also observed in irradiated GF B6. Therefore, our finding indicates that the existence of hypo-responsive subset of nTregs and that analyzing ratio of different Treg subset in SLO will be helpful to define the homeostatic status of the host. Keywords: BAFF, SOCS3, Ribosomal inactivation, Inflammation, IFN-γ Keywords: Ly6c, Natural Treg, Thymic Treg 70 The 2014 Fall Conference of the Korean Association of Immunologists Immune Mediators, Their Receptors and Signaling P-049 CCAAT-Enhancer Binding Proteinβ Expressed in Post-Switched B Cells Is Critical for Development and Maintenance of Plasma Cells GeonHee Lee, Eunkyung Jang, Jeehee Youn* Laboratory of Autoimmunology, Department of Biomedical Science, Department of Anatomy & Cell Biology, Hanyang University, Seoul, Korea Tel: 02-2290-8200, E-mail: [email protected] CCAAT/enhancer binding proteinβ (C/EPBβ), also known as nuclear factor-interleukin-6 (NF-IL6), is a basic-leucine zipper (bZIP) type transcription factor regulating survival and differentiation of diverse cells, but less was known about its role in the physiology of plasma cells. In light of our previous finding that long-lived plasma cells express C/EPBβ at the higher level than short-lived plasma cells, we hypothesized that C/EPBβ is involved in the regulatory circuit for the development and/or survival of plasma cells. To answer this hypothesis, we generated conditional knockout mice (cKO) by crossing floxed c/ebBβ mice with Cγ1-Cre mice, in which C/EPBβ gene is selectively deleted only in IgG1-switched B cells. The cKO mice and their wild-type (WT) littermates were immunized with NP-KLH/alum and assayed in terms of anti-NP antibody production. Anti-NP IgG1 titers were significantly lower in cKO mice than in WT mice, while anti-NP IgG2a and IgM titers were not different between two groups. In consistent, anti-NP IgG1 antibody-secreting cell numbers were significantly lower in both spleen and bone marrow of cKO mice than WT mice, but numbers of other isotype antibody-secreting cells were not altered. Upon stimulation with LPS and IL-4, naïve B cells sorted from cKO mice differentiated into IgG1+ memory B cells less efficiently than those from WT mice. Thus we conclude that C/EPBβ expressed in B cells at the post-switched stage is crucial for the development of plasma cells and their long lifespan in the bone marrow. Keywords: C/EBBβ, Plasma cell P-050 Poster Presentation Suppressors of Cytokine Signaling (SOCS)1 Attenuates Inflammasome Activation and Suppresses IL-1β and IL-6 Production Induced by LPS through Down-Regulation of ROS Ga-Young Kim, Hye-Young Yoon, Choong-Eun Lee* Laboratory of Immunology, Department of Biological Science, College of Natural Science, Sungkyunkwan University, Suwon, Korea Tel: 031-290-7006, E-mail: [email protected] TLR4 signaling induces a strong activation of gene expression for inflammatory cytokines. SOCS are a family of signaling molecules implicated in the regulation of inflammatory responses. Our recent finding that SOCS is induced by oxidative stress and can function in the anti-oxidant defense system prompted us to investigate the role of ROS in the regulatory action of SOCS on the inflammatory cytokine production upon LPS/TLR4 stimulation. Using SOCS-transduced and knock-down macrophage cell lines, we have observed strong inhibitory effects of SOCS1 on the production of TNF-α, IL-1β and IL-6 both at mRNA and protein levels as assessed by gene microarray analysis, qRT-PCR, and Elisa. LPS induced a transient increase in ROS levels which were suppressed upon SOCS1 transduction. The shSOCS1 cells, on the contrary, exhibited higher ROS levels and concomitant increase in the inflammatory cytokines which were both blocked by treatment of ROS scavenger NAC. The ROS-mediated inflammasome assembly was also significantly reduced by SOCS1 during TLR4 signaling with reduction in caspase-1 activation and IL-1β maturation. We have also identified the target of SOCS1 action through ROS inhibition including NF-κB and STAT1 for the LPS-induced early signaling for TNF and IL-1β gene expression. For the mechanism of ROS down-regulation by SOCS1, the role thioredoxin is investigated. In fact thioredoxin levels were up-regulated by SOCS1, and knock-down of thioredoxin gene in SOCS1-transduced cells resulted in the restoration of ROS levels and IL-6 production reduced by SOCS1. The data collectively demonstrate that potent inhibitory action of SOCS1 in inflammatory cytokine production by LPS is exerted through ROS inhibition at the level of transcription, inflammasome-mediated processing and secretion of cytokines. The induction mechanism of anti-oxidant factors with ROS-scavenging action by SOCS1 is under investigation. Supported by NRF grant #2012R1A2A2A01015258. Keywords: SOCS, TLR4, ROS, Inflammasome, Thioredoxin P-051 IFN-γ Induces Prostaglandin Production Through JAK1/2-STAT1 in HK Cell, an FDC Model Cell Jini Kim1, Jongseon Choe1,2* 1 P-052 Interleukin-32γ Increases Serum Amyloid A3 Expression in Bone Marrow-derived Dendritic Cells Jiseon Song, Tae Sung Kim* 2 Institute of Life Sciences, Kangwon National University, Department of Microbiology and Immunology, Kangwon National University School of Medicine, Chuncheon, Korea Tel: 033-250-8862, E-mail: [email protected] Department of Life Sciences, Korea University, Seoul, Korea Tel: 02-3290-3416, E-mail: [email protected] In spite of the importance of prostaglandin production by follicular dendritic cells (FDCs) in humoral immune response, cellular and molecular mechanisms to regulate prostaglandins are largely unknown. In order to investigate the effect of IFN-γ, a typical Th1 cytokine, on prostaglandin production by FDCs, we used HK cells as an FDC model. HK cells were stimulated with or without IFN-γand then the role of IFN-γin prostaglandin production was evaluated using immunoblotting, confocal microscopy, and EIA assay. IFN-γenhanced the level of COX-2, a key enzyme to produce prostaglandins at protein and mRNA level. Phosphorylation of STAT1, STAT3, and STAT5 was induced in the presence of IFNγ stimulation. However, only STAT1 siRNA diminished the effect of IFN-γ to increase COX-2 expression. IFN-γ-induced phosphorylation and the essential role of STAT1 in IFN-γ-triggered COX-2 increase depended on JAK1 and JAK2 kinases. IFN-γaugmented COX-2 protein level via STAT1 nuclear translocation and the ultimate prostaglandin production. These results indicate that IFN-γ increases prostaglandin production through JAK1/2-STAT1 pathway in HK. Our study implies that the balance between Th1 and Th2 cytokines in germinal center is critical for normal humoral immune response. Serum amyloid A3 (SAA3) is secreted during the acute phase of inflammation. SAA3 has a chemoattractant activity and induces Th17-polarizing cytokine production. Although SAA3 is induced in various cell types including LPS-treated macrophages, the expression and role of this molecule in bone marrow-derived dendritic cells (BMDCs) are still unknown. Interleukin-32γ(IL-32γ) is a pro-inflammatory cytokine, which plays a crucial part in several disease, including cancer, rheumatoid arthritis, asthma, and Crohn’s diseases. We previously reported that IL-32γinduces the expression of IL-1β, IL-12, IL-6, and TNF-αin BMDCs. In this study, we investigated whether IL-32γregulates SAA3 expression in BMDCs. Firstly, we confirmed the effect of IL-32γon the maturation of BMDCs from C57/BL6 mice. The immature BMDCs were cultured for 24h in either the absence or presence of IL-32γ. As reported, IL-32γinduced the expression of cell surface molecules, such as CD40 and CD86 as well as the mRNA expression of IL-12, IL-6, and IL-1β. Importantly, the treatment of BMDCs with IL-32γresulted in significant increase of SAA3 expression in mRNA and protein, while IL-32γtreatment didn’t affect expression of the other SAA family such as SAA1, SAA2, and SAA4. These results suggest that IL-32γmay be involved in acute phase of inflammation via increased SAA3 expression in dendritic cells. Keywords: IFN-γ, Prostaglandins, Germinal center, Humoral immune response, JAK-STAT pathway Keywords: Bone Marrow-derived Dendritic Cells, Interleukin-32γ, Serum Amyloid A3 The 2014 Fall Conference of the Korean Association of Immunologists 71 Poster Presentation P-053 Immune Mediators, Their Receptors and Signaling Anti-Inflammatory Effect of Cysteamine in Experimental Autoimmune Uveitis via Down-regulation of IL-22 Production Yejin Kim1, Tae Wan Kim2, Hyemin Kim1, Jiwon Choi1, Mirim Jang1, Jiyea Choi1, Jane Jeon1, Young Il Hwang1, Hyeong Gon Yu2, Wang Jae Lee1*, Jae Seung Kang1 1 Department of Anatomy, Seoul National University College of Medicine, 2 Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Korea Tel: 02-740-8208, E-mail: [email protected] Cysteamine modulates intracellular enzymatic activities involved in inflammation. In this study, we investigated the anti-inflammatory effects of cysteamine on experimental autoimmune uveitis (EAU) in mice. EAU was induced in female C57BL/6 wild-type mice by a footpad injection of human IRBP1-20 (250 μg/mouse) emulsified in complete Freud’s adjuvant (CFA). From one day before the IRBP inoculation, cysteamine was daily administered by intraperitoneal injection. Control group received the same amount of vehicle only. After 1 week, the draining lymph nodes were collected, and T lymphocytes were analyzed for cytokine assay by intracellular staining. In addition, when IRBP was rechallenged in vitro in splenocytes of IRBP-induced EAU mice treated with cysteamine, production of IL-22 was reduced. Both clinical and histological examinations showed that ocular inflammation was significantly delayed and decreased in cysteamine-treated mice compared to untreated mice. The amelioration of EAU in cysteamine-treated mice correlated with decreased level of IL-17-positive CD4+ T cells and IL-22Rα. The level of IL-22 was much higher in supernatant from cultured splenocytes in IRBP-induced EAU mice than in control mice and that was much lower in cysteamine-treated mice than in non-treated mice. IRBP treatment was able to stimulate the expression of IL-22Rα. But, cysteamine down-regulated the IL-22Rα expression in vivo and in vitro. We demonstrated that IL-22 can play a key role in the development of EAU. The results also show that cysteamine has an anti-inflammatory effect in EAU which may be associated with the decreased expression of Th17 cytokines and IL-22. This finding suggests that cysteamine has a beneficial effect for the control of endogenous ocular inflammatory diseases. P-054 Inhibition of IL-1beta Signaling by Zinc Leads to Repression of Memory Th17 Response in Humans Yeon ho Choi1,2, Hyunju Lee1,2, Yuri Hwang1,2, Sunjung Cho1,2, Won-Woo Lee1,2,3* 1 Department of Microbiology and Immunology, Seoul National University College of Medicine, 2Cancer Research Institute, Seoul National 3 University, Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, Korea Tel: 02-740-8303, E-mail: [email protected] Zinc is an essential trace element that plays pivotal roles in multiple facets of the immune system. Besides its catalytic and structural roles, zinc also functions as an intracellular signaling molecule, and changes in zinc levels can cause both direct and indirect modulation of immune responses. Further, cytoplasmic levels of bioavailable zinc in immune cells are largely influenced by many extracellular stimuli. Here we provide evidence + that zinc alters the cytokine production profile of human memory CD4 T cells by functioning as an intracellular signaling molecule during T-cell + responses. In vitro zinc treatment of CD4 T cells in the presence of activated monocytes inhibited IFN-γ- and IL-17-producing cells, but not IL-4-producing cells. Of note, production of IL-17+ cells from memory + CD4 T cells, which is significantly up-regulated by LPS-stimulated monocytes, was preferentially repressed by zinc. Increased cytoplasmic zinc in CD4+ T cells suppressed IL-1β signaling via repression of phosphorylation of IRAK4, thus leading to an inhibitory effect on Th17 responses facilitated by monocyte-derived IL-1βin humans. These finding suggest that therapeutic manipulation of zinc bioavailability may be a + good means by which to modulate memory CD4 T cell responses. Keywords: Zinc, Th17 cells, Monocytes, IL-1β, IRAK4 Keywords: Cysteamine, Experimental autoimmune uveitis, Interleukin-22 P-055 Imaging of Cytotoxic Dynamics of CXCR3-Deficient Cytokine-Induced Killer Cells at the Single Cell Level Yong Guk Kim, Ji Sung Kim, Juyoung Kim, Eun Jae Park, Hong Kyung Lee, Hwa Sun Ryu, Minji Pyo, Jin Bum Uhm, Jin Tae Hong, Youngsoo Kim, Sang-Bae Han* College of Pharmacy, Chungbuk National University, Cheongju, Korea Tel: 043-261-2815, E-mail: [email protected] Cytokine-induced killer (CIK) cells are a population of cytotoxic T cells having characteristics of natural killer cells. CIK cell-mediated cytotoxicity is mainly controlled by direct cell-cell interaction and usually studied in population level. However, little is known about cytotoxic dynamics of CIK cells at the single cell level. Using time-lapse imaging, we examine how CIK cells migrate, contact, and kill cancer cells in vitro and whether cancer cell-derived chemokines affect this response. B16F10 melanoma target cells highly express CXCL10, which could bind +/+ CXCR3 expressed in CIK cells. During migration stage, CXCR3 CIK cells moved at a speed of 6.3 μm/min and CXCR3−/− CIK cells moved at a speed of 5.1 μm/min, which suggested that cancer cell-derived +/+ CXCL10 might increase chemokinesis of CXCR3 CIK cells. Of interest was both CIK cells showed random migration, but not directed ones. During contact stage, CXCR3+/+ contacted melanoma cells for 31 min per −/− each contact and CXCR3 CIK cells did for 19 min. During cytotoxic stage, CXCR3−/− CIK cells killed much fewer target cells than CXCR3+/+ CIK cells did, which might be a consequence of defective migration and contact. Together, our data suggest that cancer cell-derived CXCL10 regulates cytotoxic dynamics of CIK cells: increase of the migration speeds, contact ability, and cytotoxicity of CIK cells. However, CIK cells migrate randomly, not directionally, in culture condition having cancer-derived chemokines. Keywords: CIK (Cytokine-Induced killer cells), Dynamics, Melanoma, Chemokinesis, CXCR3 P-056 N-linked Glycosylation of HMGB1 Plays Critical Role in Its Secretion Young Hun Kim1,2, Man Sup Kwak1, Jun Bae Park3, Shin-Ae Lee1, Hyun-Soo Cho3, Jeon-Soo Shin1,2* 1 2 Department of Microbiology, Severance Biomedical Science Institute and Institute for Immunology and Immunological Diseases, Yonsei University College of Medicine, 3Department of Systems Biology, Yonsei University College of Life Science and Biotechnology, Seoul, Korea Tel: 02-2228-0800, E-mail: [email protected] HMGB1 is a nuclear protein, known as a delayed mediator of sepsis, secreted to the extracellular milieu as a response to various stimulants which then stimulates cytokine production as a DAMP molecule. HMGB1 is secreted after being subjected to various post-translational modifications such as acetylation, phosphorylation, and methylation. Here, we identified a novel role of N-linked glycosylation in HMGB1 secretion. First of all, we observed glycosylation of recombinant HMGB1 using a glycoprotein staining and disappearance of glycosylation by PNGase F treatment. And we have observed N-linked glycosylation of endogenous HMGB1 through HMGB1 band shift-down after treatment with N-linked glycosylation inhibitor tunicamycin in CHO, HeLa, RAW264.7 cell lines. In MALDI-ToF analysis, N37, N134, and N135 residues were revealed to be the sites for N-linked glycosylation. We constructed N-linked glycosylation mutants by mutating Asp to Gln, and investigated the secretion of HMGB1. HEK293T cells transfected with HMGB1 mutant plasmids of N37/134Q and N37/135Q showed dramatically decreased extracellular secretion of HMGB1, low cytoplasmic localization, and faster protein degradation after treating with various stimulants. The binding affinity of HMGB1 mutant protein to nuclear DNA and nuclear export protein CRM1 showed stronger binding to nuclear DNA but lower binding to CRM1, respectively. These data suggest that HMGB1 is N-glycosylated and essential for the secretion of HMGB1. This research was supported by grants from the National Research Foundation of Korea funded by the Korea government (MEST) (No. 20110017611 and 2014R1A4A1008625) and the Brain Korea 21 PLUS Project for Medical Science, Yonsei University. Keywords: DAMP molecule, HMGB1, N-linked glycosylation, Secretion 72 The 2014 Fall Conference of the Korean Association of Immunologists Immune Mediators, Their Receptors and Signaling P-057 Mast Cell 2B4 Regulates Activation of Antigen-Specific CD8+ T Cells Jiyoung Kim, Kwanghee Kim, Junho Lee, Gayoung Park, Seon Ah Lim, Kyung-Mi Lee* Department of Biochemistry and Molecular Biology, Korea University, Seoul, Korea Tel: 02-920-6251, E-mail: [email protected] 2B4 (CD244), belonging to signaling lymphocyte activation molecule (SLAM) receptor family, is expressed on NK cells, monocytes, γδ-T cells, and dendritic cells. Recently 2B4 was also shown to be expressed on mast cells (MCs) derived from bone marrow, however the role of 2B4 in mast cells is not known. More recent studies suggest that MCs could function as APCs and regulate CD8+ T cell effector functions both in vivo and in vitro. Since the ligand of 2B4, CD48, is ubiquitously expressed on CD8+ T cells, we hypothesized that 2B4 expressed on mast cells could function to costimulate antigen-specific T cells via binding to CD48. To address this question, we co-cultured isolated OT-I CD8+ T cells from the spleen with OVA257-267 (SIINFEKL)-pulsed wild type (WT) or 2B4 KO mouse bone-marrow derived MCs (BMMCs) and examined their activation status. Here we show that CD8+ T cells cultured with OVA pulsed WT BMMCs demonstrated higher expression of CD69 and CD25 activation markers but not 2B4 KO BMMCs. To confirm whether MC induced CD8+ T cells activation depends on 2B4 molecule, we blocked several molecules on MCs using each blocking antibodies. The blocking effect of 2B4 reduced activation of CD8+ T cell, but also CD48 on MC was involved regulation of CD8+ T cell. These data demonstrate that 2B4 can function, not only as a receptor, but also as a ligand to influence T antigen-specific CD8+ T cell responses. Keywords: 2B4, CD48, Mast cell, Antigen presenting cell, CD8+ T cell P-058 Poster Presentation The Xanthine Oxidase-NFAT5 Pathway Regulates Macrophage Activation and TLR-Induced Inflammatory Arthritis Susanna Choi1†, Nam-Hoon Kim1†, Eun-Jin Han1, Bong-Ki Hong1, Soo Youn Choi2, H Moo Kwon2, Sue-Yun Hwang3, Chul-Soo Cho1,4, Wan-Uk Kim1,4* 1 POSTECH-CATHOLIC BioMedical Engineering Institute, The Catholic University of Korea, Seoul, 2School of Nano-Bioscience and Chemical Engineering, Ulsan National Institute of Science and Technology, Ulsan, 3 Department of Animal Biotechnology, Graduate School of Biology and Information Technology, Hankyoung National University, Ansung, 4Department of Internal Medicine, Catholic University of Korea, Seoul, Korea † Susanna Choi and Nam-Hoon Kim contributed equally to this work. Tel: 02-2258-7530, E-mail: [email protected] NFAT5, a well known osmo-protective factor, can be activated by isotonic stimuli, such as Toll-like receptor (TLR) triggering. However, it is unclear how NFAT5 discriminates between isotonic and hypertonic stimuli to produce different functional and molecular outcomes. Here, we identified a novel xanthine oxidase (XO)-reactive oxygen species (ROS)-p38 mitogen-activated protein kinase-NFAT5 pathway that is activated in RAW 264.7 macrophages upon isotonic TLR stimulation. Unlike what is seen under hypertonic conditions, XO-derived ROS were selectively required for the TLR-induced (isotonic) NFAT5 activation and NFAT5 binding to the IL-6 promoter in RAW 264.7 macrophages. In mouse peritoneal macrophages and human macrophages, TLR ligation also induced NFAT5 activation, which was dependent on XO and p38 kinase. The involvement of XO in NFAT5 activation by TLR was confirmed in RAW 264.7 macrophages implanted in BALB/c mice. Moreover, allopurinol, a XO inhibitor, suppressed arthritis severity in C57BL/6 mice and decreased the expression of NFAT5 and IL-6 in splenic macrophages of the mice. Collectively, these data support a novel function of the XO-NFAT5 axis in macrophage activation and TLR-induced arthritis, and suggest that XO inhibitor(s) could serve as a therapeutic agent for chronic inflammatory arthritis. Keywords: Xanthine oxidase, Reactive oxygen species, NFAT5, Innate immunity, Inflammatory arthritis P-059 GREM1 is a Key Regulator of Synovial Hyperplasia in Rheumatoid Arthritis Eun-Jin Han1, Daehee Hwang2, Sungyong You3, Wan-Uk Kim1,4* 1 POSTECH-CATHOLIC BioMedical Engineering Institute, The Catholic University of Korea, Seoul, 2Center for Systems Biology of Plant Senescence and Life History, Institute for Basic Science, DGIST, Daegu, 3 Korea, Departments of Surgery and Biomedical Sciences, Cancer Biology Program, Smuel Oschin Comprehensive Cncer Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA, 4Department of Internal Medicine, The Catholic University of Korea, Seoul, Korea Tel: 02-2258-7530, E-mail: [email protected] Rheumatoid arthritis (RA) is an autoimmune disease leading to chronic inflammation of the joints orchestrated by various inflammatory cells in the synovium. Fibroblast like synoviocyte (FLS) plays critical roles is the major cell in the inflammatory processes of RA. However, understanding molecular mechanisms underlying aberrant activation of RA FLS have not been fully elucidated. Thus, we attempted to identify a key regulator that can drive aggressive phenotypes of RA FLS. For this, we performed an integrative transcriptome data analysis using three datasets from independent RA studies, a panel of normal tissues, and NCI60 cancer cell line panel. Through the analysis, we identified Gremlin (GREM1), which is robustly up-regulated in RA and predominantly expressed in FLS, as a novel candidate of FLS activation. GREM1 is originally identified as bone morphogenetic proteins (BMP2, BMP4 and BMP7) antagonist. To investigate potential roles of GREM1 in RA context, we first examined the level of GREM1 expression in synovial tissues, synovial fluid and FLS from RA and ones from osteoarthritis (OA). Immunohistochemistry and ELISA revealed up-regulation of GREM1 protein in RA compared to OA. Proinflammatory cytokines, such as IL-1β, TGF-β, and TNF-α, also induced the expression of GREM1 in RA FLS and their expressions were significantly correlated with GREM1 expression in RA SF. We next investigated cellular functions of GREM1 using RA FLS. GREM1 deficiency by siRNA elicited significant suppression of FLS proliferation, migration and invasion. In contrast, stimulation with rhGREM1 to RA FLS increased cell survival and invasion. Furthermore, we found that GREM1 activates AKT1 and MAPK signaling cascade, as well as antagonizes BMP2. This study demonstrated that GREM1 is a key regulator responsible for synovial hyperplasia and FLS migration through activation of AKT and MAPK signaling cascades and inhibition of BMP2 mediated synoviocytes apoptosis in rheumatoid synoviums. P-060 Leukotrienes Act as Chemoattractant to Th17 Cells Hyeong-Su Kim, Wonyong Lee, Gap-Ryol Lee* Department of Life Science, Sogang University, Seoul, Korea Tel: 02-705-8458, E-mail: [email protected] Th17 cells are one of CD4+ T cells that have crucial role in inflammation and autoimmunity. Although many studies revealed molecular mechanisms about Th17 cells but the traffic of Th17 cells by lipid-based chemoattractant leukotriene signaling is not fully understood. Here, we show that Th17 cells are highly sensitive to leukotriene B4 and D4. Th17 cells expressed high levels of leukotriene B4 receptors and cysteinyl leukotriene receptors compare to other effector CD4+ T cell subsets. In chemotaxis assay, we confirmed that migration of Th17 cells was more efficient than that of other effector T CD4+ T cell subsets under the guidance of leukotriene B4 and D4, and it was blocked by specific inhibitors of leukotriene B4 receptor and cysteinyl leukotriene receptor. Finally in an animal disease model of experimental autoimmune encephalomyelitis (EAE), treatment with montelukast which is an inhibitor of cysteinyl leukotriene receptor, it mitigated clinical score of EAE. Thus, we concluded that leukotrienes act as chemoattractant for Th17 cells. Keywords: Th17, LTB4, LTD4, Montelukast, EAE Keywords: Gremlin, Bone morphorgenetic protein 2, Fibroblast like synoviocyte, Rheumatoid arthritis The 2014 Fall Conference of the Korean Association of Immunologists 73 Poster Presentation P-061 Immune Response Regulation and Tolerance The Discovery and Validation of Disease Specific Proteins in CRH-Treated Regulatory T Cells in Atopic Dermatitis Shan Jin1,2, Jung U Shin1, Ji Yoen Noh1, Chang Ook Park1, SeoHyeong Kim1,2, Hemin Lee1, Jungsoo Lee1, Kwang Hoon Lee1,2* 1 Department of Dermatology & Cutaneous Biology Research Institute, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea Tel: 02-2228-2084, E-mail: [email protected] 2 Most of atopic dermatitis (AD) patients were triggered or exacerbated by psychological factors, such as stress. Stress induces corticotropin-releasing hormone (CRH) in the CNS as well as in peripheral sites. Regulatory T cells (Tregs) are a subset of T cells with strong immunosuppressive activity. However, the role of Tregs in AD with stress has not been investigated. Therefore, the aim of this study was to discover CRH-regulated protein and its role in AD Tregs. Female NC/Nga mice were applied with Dermatophagoides farinae body extract (DfE) ointment for six weeks. After sensitizing NC/Nga mice for six weeks, we observed AD-like + + skin lesions, elevated serum IgE levels, increased CD4 IL-4 Th2 cells and in+ + creased CD4 CD25 Tregs. Splenocytes from AD mice were treated with 10 nM + + CRH for 24 and 48 hours, and then CD4 CD25 Tregs were sorted. Differential protein expression between CRH-treated group and non-treated group was compared by Tandem Mass Tag (TMT) labeling method. Initially, we identified 1353 proteins in CRH-treated and non-treated Tregs. Afterwards, we attempted to identify proteins that either increased or decreased depending on time after stimulation with CRH. Finally, we identified 20 significantly up-regulated (5 proteins) or down-regulated (15 proteins) proteins. Among the down-regulated proteins, we paid special attention to DOCK8 because of its role in various immune diseases. Using Western blot and flow cytometry analysis, we confirmed decrease of DOCK8 in CRH-treated Tregs in AD mouse and AD patients. We finally found that inhibitory cytokine TGF-βand IL-10 were decreased in DOCK8 siRNA-treated Tregs. In summary, CRH-induced DOCK8 down-regulation might result in the inhibition of TGF-βand IL-10 expression, which may indicate the impairment of Treg function. In conclusion, we suggest that DOCK8 may play an important role in Tregs which may contribute to alleviating stress induced aggravation of AD symptom. Keywords: Atopic dermatitis, Regulatory T cell, Corticotropin-releasing hormone, Dedicator of cytokinesis 8 P-063 Department of Physiology, College of Korean Medicine, Kyung Hee University, Seoul, Korea Tel: 02-961-9316, E-mail: [email protected] Bee venom (BV) is one of the most traditional medicines that have been widely used in the treatment of chronic inflammatory diseases, such as rheumatoid arthritis. In the previous study, we reported that BV induced immune tolerance by increasing the population of regulatory T cells (Tregs) in lupus nephritis, cisplatin-induced nephrotoxicity and allergic asthma model. But what component and how it regulates the immune response is mostly unclear. Here, we investigated whether bee venom phospholipase A2 (bvPLA2) has protective effects that are mediated via Tregs in OVA-induced allergic asthma model. We found that bvPLA2 treatment increased the Treg population and suppressed the production of Th2 cytokines in the airways of OVA-induced asthma mice. To assess the involvement of Treg in the bvPLA2 effects, Treg were depleted by anti-CD25 mAb. Although bvPLA2 significantly reduced the number of inflammatory cells in the lung parenchyma and the BAL fluid after OVA challenge, these effects were not observed in Treg-depleted mice. In addition, CD206-deficiency showed the bvPLA2-mediated attenuation of airway inflammation depended on the binding to its receptor, macrophage mannose receptor (CD206). Our results strongly suggest that bvPLA2 has potential as an effective agent on Treg recruitment into the airway for controlling allergic asthma. This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korean government (Ministry of Education, Science, and Technology, MEST) (no. 2007-0054934). Keywords: Bee Venom, Regulatory T cells, Allergic asthma, CD206 L. helveticus Suppresses experimental rheumatoid arthritis by Reducing Inflammatory T Cell Responses Jung-Eun Kim1,2,3, Chang Suk Chae2, Gi-Cheon Kim1,2,3, 1,2,3 2,3 2,3 Won Hwang , Changhon Lee , Sung-Min Hwang , 4 5,6 7 Young-Tae Ahn , Chul-Sung Huh , Young Kim , So-Young Lee8, Sung-Gyoo Park1, Chang-Duk Jun1, 2,3 Sin-Hyeog Im * 1 School of Life Sciences, Gwangju Institute of Science and Technology (GIST), Gwangju, 2Academy of Immunology and Microbiology (AIM), 3 Institute for Basic Science (IBS), Division of Integrative Biosciences and Biotechnology (IBB), Pohang University of Science and Technology, Pohang, 4R&BD Center, Korea Yakult Co. Ltd., Yongin, 5Department of Food and Animal Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul, 6Research Institute of Eco-friendly Animal Sciences, Institute of Green Bio Science & Technology, Seoul 7 National University, Pyeongchang, Chonnam National University Medical School, Gwangju, 8Fermentation Research Center, Korea Food Research Institute, Sungnam, Korea Tel: 054-279-2356, E-mail: [email protected] Probiotics confer a health benefit when administered in adequate amount to modulate diverse pathophysiological conditions. However efficacy of probiotics is strain specific and differs within individuals. Identification of functional probiotics to modulate a pathophysiological condition is a challenging task. To identify the anti-inflammatory probiotics among the diverse probiotic strains we performed an ex vivo screening that mimics the gut associated lymphoid tissue. Probiotic strains were co-cultured with mesenteric lymph node cells, and candidate strains that induce IL-10highIL-12low levels were selected. Three candidate strains were selected based on their high (>10) IL-10/IL-12 ratio without showing no cytotoxicity in the co-culture system. Among them, we selected a Lactobacillus helveticus HY7801 as the best candidate based on its prophylactic effects on suppression of the experimental rheumatoid arthritis. Oral administration of L. helveticus suppressed inflammatory arthritis by reducing a disease incidence, antigen specific IgG levels, and an infiltration of immune cells into the synovial region. Administration of L. helveticus also altered cytokine production profile by reducing the levels of pro-inflammatory cytokines (TNF-α, IFN-γ, and IL-17A) while enhancing anti-inflammatory cytokine IL-10. This phenotypic changes induced by L. helveticus was mediated by CD11c+ dendritic cells in mesenteric lymph node, which confer CD4 T cells to produce high levels of IL-10. Our study suggest that screening of probiotic strains based on the IL-10highIL-12low selection criteria may applicable to identify anti-inflammatory probiotics for the modulation of inflammatory immune responses including rheumatoid arthritics. This research was supported by Global PH.D Fellowship Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, and by research grants from the Korea Food Research Institute and from a research program for Agricultural Science & Technology Development (Project No: PJ907153), National Academy of Agricultural Science, Rural Development Administration, Republic of Korea. Keywords: Probiotics, Immune modulation, Rheumatoid arthritis, Dendritic cells, T helper cells Bee Venom Phospholipase A2 Suppresses Airway Inflammation by Inducing Regulatory T Cells in an OVA-Induced Asthma Model Hyunjung Baek, Soojin Park, Kyung-Hwa Jung, Dasom Shin, Gihyun Lee, Hyeonhoon Lee, Geun-Hyung Kang, Hyunsu Bae* 74 P-062 P-064 Progressive Epigenetic Regulation of IL-10 Gene Expression during Th1 and Th2 Differentiation Changhon Lee1,2, Won Hwang1,2,3, Sin-Hyeog Im1,2* 1 Academy of Immunology and Microbiology (AIM), Institute for Basic Science (IBS), Pohang, 2Division of Integrative Biosciences and Biotechnology (IBB), Pohang University of Science and Technology, 3 Pohang, School of Life Sciences, Gwangju Institute of Science and Technology (GIST), Gwangju, Korea Tel: 054-279-2356, E-mail: [email protected] The immunoregulatory cytokine IL-10 has many functions especially in maintaining the balance between immunity and tolerance. IL-10 is expressed mainly by T helper 2(Th2) cells but also by T helper 1(Th1) cells during chronic infection. Although IL-10 is produced by both in vitro primary Th1 and Th2 polarizing conditions, Th2 cells produce much higher levels than Th1 cells. However, little information is available on the epigenetic mechanisms of IL-10 gene regulation at the transcriptional level. In this study, we have investigated the role of epigenetic modification on IL-10 gene regulation by testing main epigenetic marker such as DNA modification around IL-10 regulatory elements. To elucidate long term epigenetic memory, we gave repetitive Th1 and Th2 polarizing conditions from round1(R1) to round3(R3) to mouse primary CD4+ T cells respectively. Restriction enzyme cleavage assay, pyrosequencing analysis and methylated DNA immunoprecipitation were performed to measure the DNA methylation statues in Th1 and Th2 cells from R1 to R3. Indeed, methylated DNA levels were much higher in Th1 cells, which are well correlated with lower IL-10 gene expression on Th1 cells than Th2 cells. Especially methylated DNA levels were increased in Th1 cells during repetitive Th1 and Th2 skewing conditions In vitro DNA methylation on CpG rich regions of IL-10 gene locus significantly reduced IL-10 promoter activity. Treatment of 5-Azacytidine, DNA methyltransferase inhibitor, significantly up-regulated IL-10 mRNA expression in Th1 cells. The strongest effect of 5-Azacytidine was shown in R1 Th1 cell as compare to R2, R3 Th1 cell. To check the ability of plasticity, Th2 polarizing conditions were given to skewed Th1 cell from R2 to R3 respectively. However, IL-10 expression level and DNA methylation status were not changed in this system. It is reported that IL-12 is one of the positive signaling molecules for IL-10 expression. Based on this, we kept the concentration of IL-12 during Th1 differentiation in vitro. IL-10 mRNA expression was increased 2-fold by keeping IL-12 concentration. However, pyrosequencing analysis revealed that DNA methylation profile in the IL-10 locus was not changed. Collectively, those results suggest that DNA modification around IL-10 regulatory elements play crucial role for differential IL-10 gene expression between Th1 and Th2 cells. Those DNA methylation profile in Th1 cells are not flexible under Th2 cell differentiation condition in vitro. Under keeping IL-12 concentration during Th1 cell differentiation, which mimicking chronic inflammation condition, IL-10 expression is up-regulated but DNA methylation profile was not changed. Based on those results, DNA methylation status was well correlated to IL-10 expression during common Th1 and Th2 cell differentiation but not related with flexibility in Th2 differentiation conditions and chronic inflammation conditions. This work was supported by IBS-R005RG1-2014-a00 and by research grants from the Korea Food Research Institute. Keywords: IL-10, DNA methylation, T helper 1 cell, T helper 2 cell The 2014 Fall Conference of the Korean Association of Immunologists Immune Response Regulation and Tolerance P-065 Interleukin-18 Enhances Immunosuppressive Responses by Promoting Differentiation into Monocytic Myeloid-Derived Suppressor Cells Hui Xuan Lim1, Hye-Jin Hong1, Daeho Cho2, Tae Sung Kim* 1 Division of Life Sciences, College of Life Sciences and Biotechnology, Korea University, 2Department of Life Science, Sookmyung Women’s University, Seoul, Korea Tel: 02-3290-3416, E-mail: [email protected] Myeloid-derived suppressor cells (MDSCs) are major immunosuppressive cells that lead to T cell defects in cancer. Interleukin-18 (IL-18) is important in inflammatory and immune responses. IL-18 has been reported to have a dual effect on tumor progression, as it not only stimulates host immune responses, but also exerts pro-cancer effects by inducing immune escape and angiogenesis. In the present study, we investigated the effect of IL-18 on MDSCs and found that IL-18 treatment significantly increased the percentage and the absolute number of monocytic MDSCs − (M-MDSCs) via differentiation of CD11b bone marrow (BM) progenitor cells. IL-18-induced MDSCs showed enhanced suppression of T cell proliferation and IFN-γ production along with a dramatic increase of M-MDSC suppressive function including nitric oxide production and arginase 1 expression. Although IL-18 decreased the number of granulocytic MDSCs (G-MDSCs) in a concentration-dependent manner, we found that the absolute number of G-MDSCs and their reactive oxygen species production remained unchanged. Additionally, we demonstrated that IL-18-induced M-MDSCs have a more potent suppressive effect on T cell responses with lower IFN-γproduction than G-MDSCs, suggesting that the increased suppressive effect observed in our study was resulted from M-MDSCs. Furthermore, in vivo administration of IL-18 significantly increased the accumulation of M-MDSCs in tumor microenvironment. Taken together, our findings indicate that IL-18 specifically enhances the differentiation and function of M-MDSCs, leading to immunosuppression. P-066 Poster Presentation Time-Lapse Imaging of Dynamic Interaction between Mesenchymal Stem Cells and T Cells Hong Kyung Lee, Hwa Sun Ryu, Minji Pyo, JinBeom Eom, Ji Sung Kim, Yong Guk Kim, Juyoung Kim, Eun Jae Park, Jin Tae Hong, Youngsoo Kim, Sang-Bae Han* College of Pharmacy, Chungbuk National University, Cheongju, Korea Tel: 043-261-2815, E-mail: [email protected] Mesenchymal stem cells (MSC) ameliorate the pathogenesis of systemic lupus erythematosus through inhibiting T cell functions via soluble mediators and direct cell-cell contact. However, little is known about the contact dynamics at the single cell level. Using time-lapse imaging, we show that their contact is dependent on CCL2, but not CCL3, CCL4, CXCL10, and CXCL12, which are produced by MSCs. First, we analyzed contact dynamics of MSCs. Each WT MSC contacted 11 T cells and each contact lasted 107 min. In the same condition, each CCL2−/− MSC contacted 10 −/− MSCs T cells and each contact continues 34 min. Subsequently, CCL2 did not inhibit T cell functions. Next, we analyzed migration dynamics of only T cells, since MSCs did not move well. When calculating the percentage of T cells contacting MSCs more than one time for 6 h, 68% T −/− cells contacted MSCs and 59% T cells contacted CCL2 MSCs. T cells moved at average speeds of 7 um/min without contact and did not move well during the contact period, which was similar in cultures with either −/− WT or CCL2 MSCs. Together, our data demonstrate that MSC-derived CCL2 might increase T cell migration, prolong contact duration with T cells, and finally inhibit T cell functions. Keywords: Mesenchymal stem cells, CCL2 Keywords: IL-18, Myeloid-derived suppressor cells, Immunosuppression, NO, Tumor P-067 Glucans Isolated from Morus alba Enhances Lymph Node Homing of Dendritic Cells through Increasing CCR7 Expression Hwa Sun Ryu, Mi Jeong Park, Yong Guk Kim, Ji Sung Kim, Juyoung Kim, Eun Jae Park, Hong Kyung Lee, Minji Pyo, Jin Tae Hong, Youngsoo Kim, Sang-Bae Han* College of Pharmacy, Chungbuk National University, Cheongju, Korea Tel: 043-261-2815, E-mail: [email protected] Maturation and activation of dendritic cells (DC) are critical factors for initiating the innate and adaptive immune response. However, DC functions are usually attenuated in the tumor microenvironment, which is an important immunological problem in DC-based immunotherapy against cancer. In this study, we report the effect of Morus alba extracts (MAE) on DC functions. MAE increased CD40, CD80, CD86, and major histocompatibility complex-I/II expression in DC surface, showing the phenotypic maturation of DC. MAE induced the functional activation of DCs, in that MAE increased the production of IL-12, IL-1β, TNF-α, and IFN-γ, and deceased antigen capture capacity of DCs. MAE enhanced allogeneic T cell activation ability of DCs. MAE increased the expression of CCR7, but decreased the expression of CCR1, CCR2, and CCR5. Subsequently, MAE increased DC migration to MIP-3βin in vitro transwell assay and lymph node (LN) homing when injected into footpads of mice. Knockdown of CCR7 with siRNA decreased the in vitro migration and in vivo LN homing ability of MAE-treated DCs. In summary, MAE induces the maturation and functions of DCs and enhances LN homing of DCs through increasing CCR7 expression. P-068 Saucerneol D Inhibits Dendritic Cell Functions by Heme Oxygenase-1 Induction Hwa Sun Ryu, Hong Kyung Lee, Minji Pyo, JinBeom Eom, Ji Sung Kim, Yong Guk Kim, Juyoung Kim, Eun Jae Park, Jin Tae Hong, Youngsoo Kim, Sang-Bae Han* College of Pharmacy, Chungbuk National University, Cheongju, Korea Tel: 043-261-2815, E-mail: [email protected] Saucerneol D (SD) isolated from the roots of Saururus chinensis has been reported to have antioxidant, anti-asthmatic, and anti-inflammatory activities, but its mechanisms of action remain to be defined. In this study, we examine the inhibitory mechanisms of SD on dendritic cells (DCs), which play important role in innate and adaptive immunity. In phenotype assay, SD decreased the expression of MHC and co-stimulatory molecules in LPS-treated dendritic cells. In functional assay, SD also decreased NO production, antigen capture capacity, pro-inflammatory cytokine gene expression, and allogenic T cell activation capacity of LPS-treated DCs without cytotoxicity. In molecular assay, SD attenuated NF-κB and MAPK signaling, not TRIF signaling in LPS-treated DCs, whereas SD did not affect directly IRAK4, IRAK1, TAK1 and IKKβkinase activity in enzymatic assays. Finally, we proved that SD increased the expression of heme oxygenase (HO)-1 via mediating ROS-Nrf-2 activation in DCs, which resulted in overall suppression of DC functions. Together, our data suggest that SD can inhibit DC functions through inducing HO-1, but not directly inhibiting NF-kB and MAPK signaling molecules. Keywords: Saucerneol D, TLR4, Heme oxygenase-1 Keywords: Morus alba extract, Dendritic cell, CCR7, LN homing The 2014 Fall Conference of the Korean Association of Immunologists 75 Poster Presentation P-069 Immune Response Regulation and Tolerance IL-10-Producing Regulatory B Cells in Neuromyelitis Optica Spectrum Disorder and Multiple Sclerosis P-070 SOCS-1 Triggers Mouse Macrophage Tolerance through the Limited Activation of NF-κB Signaling in TLR Pathway Hye-Jin Cho, Byoung Joon Kim, EunJu-Hong Min* Hyo-Ji Lee1,2, Yu-Jin Jung1,2* Department of Neurology, Samsung Medical Center, Sungkyunkwan University, Korea Tel: 02-3410-2725, E-mail: [email protected] 1 Whereas the main pathogenesis of multiple sclerosis(MS) is considered to be mediated by Th1, that of neuromyelitis optica(NMO) may be humoral immune mechanism associated with aquaporin 4-antibody(AQP4-ab). But still many aspects of the pathogenic cascade in NMO and MS remain to be determined especially in the lymphocyte subsets. We investigated the balance of the proinflammatory lymphocytes and suppressive lymphocytes comparing both MS and NMO to the healthy subjects. Twenty-three seropositive NMO patients, eleven MS patients, and 13 healthy controls were enrolled in this study. The skewness of B cell, regulatory B (Breg) cell,IL-10 secreting Breg(B10), T cell, T helper (Th) cell, cytotoxic T (Tc) cell, Th1 cell, Th2 cell, Th17 cell, and regulatory T(Treg) cell among lymphocytes were measured by flow cytometry. The proportion of B cells was higher in the MS group than in the NMOSD group or HC (p=0.003) and the proportion of B10 was significantly higher in both NMOSD and MS group than HC (p<0.001). Only the proportion of B10 showed significant correlation with AQP4-ab intensity (p=0.02, rho=0.40). The proportion of B cells was significantly higher in the attack state than in the remission state in seropositive NMOSD group. (p<0.001) and the proportion of Tc was significantly lower in the attack state. Our results suggest that the elevated B cells in MS in both remission and relapse state could be the differential marker of these two diseases. And the elevation of B cell of NMO patient may be the indicator of the relapsing. The elevation of B10, Tc, and Th2 cells in NMO patients and elevation of B10 cell in MS patients may be due to the immunomodulating drug like azathioprine or compensatory increasing of dysfunctional cells. B1B10 is the unique subset that showed B10 is the unique subset that showed correlation with the AQP4-ab intensity, so, the B10 would be the sensitive and important compensatory mechanism in NMO patient. Toll-like receptors (TLRs) are involved in host defense to recognize infectious pathogens and to activate innate immune system. Sensing of microbial components by TLRs triggers the activation of NF-κB or MAPK signaling to induce the expression of pro-inflammatory cytokine genes, such as TNF-α, IL-6 and IL-12. As known as that excessive TLR activation can induce variety of severe inflammatory response, TLR signaling must be tightly regulated. In this view, several negative regulators of TLR signaling, such as IRAK-M, SOCS-1 and A20, have been studied well. In this study, it was verified which kind of negative regulators could be involved in macrophages in excessive stimulatory conditions through endosomal TLRs. When macrophages were simultaneously stimulated with both TLR7 agonist, Gardiquimod (GDQ), and TLR9 agonist, CpG ODN 1826, co-treatment of both TLR agonists lowered the production of TNF-αand IL-6 through the delayed activation of NF-κB pathway, as compared with individually stimulated cells. The decreased nuclear translocation of NF-κB p65 was also observed in co-stimulated macrophages. In addition, simultaneous stimulation of both TLR agonists induced the increased level of SOCS-1. The specific knockdown of SOCS-1 using siRNA was able to rescue the secretion of TNF-α and NF-κB activity in co-stimulated mouse macrophages, which coincided with kinetics of degradation and phosphorylation of IκBα. Following co-stimulation of TLR7 and TLR9, SOCS-1 and NF-κB p65 subunit translocated to the nucleus, where NF-κB p65 interacted with SOCS-1 and was markedly ubiquitinated. Moreover, the expression of TLR7 and TLR9 was reduced at both transcriptionally and translationally when cells were treated simultaneously with the agonists. These findings revealed that co-stimulation and/or re-stimulation of TLR7 and TLR9 induced macrophage tolerance with decreased expression of inflammatory cytokines through the inhibitory function of SOCS-1 to NF-κB signaling pathway. Keywords: Neuromyelitis optica, Multiple sclerosis, Anti-aquaporin4-antibody, Regulatory B cell, Cytotoxic T cell Keywords: Macrophage, NF-κB, TLRs, SOCS-1, Tolerance P-071 BIT Medical Convergence Graduate Program, 2Department of Biological Sciences, Kangwon National University, Chuncheon, Korea Tel: 033-250-8533, E-mail: [email protected] An Acidic Polysaccharide of Panax ginseng Rescues Irradiated Mice by Accelerating Hematopoiesis and Curtailing Apoptosis Jinhee Cho1, So Jin Bing1, Eunjin Park1, Dae Seung Kim1, 1 2 1 Areum Kim , Jie-Young Song , Youngheun Jee * 1 Department of Veterinary Medicine and Institute for Nuclear Science and 2 Technology, Jeju National University, Jeju, Laboratory of Radiation Immunology, Korea Institute of Radiological and Medical Sciences, Seoul, Korea Tel: 064-754-3374, E-mail: [email protected] The radiation therapy is a common choice of treatment for various diseases however surrounding normal tissues are also affected. It is essential to identify efficient, safe and affordable radio-protectors in radiosensitive tissue such as hematopoietic cells and gastrointestinal tract. However, synthetic chemicals have severe side effects such as vomiting, nausea, diarrhea, and neurotoxicity at clinically effective doses. An acidic polysaccharide isolated from Panax ginseng (APG) has been reported to stimulate the proliferation of normal lymphoid cells and produce cytokines such as IL-1, IL-2, IFN-γand TNF-α. Here, we evaluated the radio-protective effects of APG in immune system and small intestines and investigated its protective mechanism in detail. APG (10mg/kg b.w.) was injected intraperitoneally twice into mice, first at 18 h and then again at 2h, before irradiation. The counts of endogenous spleen CFUs increased in APG-treated mice, indicating that APG induced the regeneration of hematopoietic cells. APG not only minimized DNA damages of peripheral immunocytes but also stimulated cell proliferation of lymphocyte. Furthermore, the number of apoptotic cells was significantly decreased in lymphocytes of APG-treated mice. In the small intestines, APG treatment promoted the lengthening of villi and the regeneration of crypts. APG pretreatment dramatically decreased the radiation-induced apoptosis in jejunum. The expression level of anti-apoptotic (Bcl-2 and Bcl-XS/L) proteins was markedly increased whereas that of pro-apoptotic (p53, BAX, cytochrome-C and Caspase-3) proteins was significantly reduced. These results suggest that APG can be a candidate for a novel non-toxic radio-protector. This work was supported by the project titled 2014 Jeju Sea Grant funded by the Ministry of Land, Transport and Maritime Affairs of Korea and High Value-added Food Technology Development Program, Ministry of Agriculture, Food and Rural Affairs. P-072 Nontuberculous Mycobacterial Lung Disease Indicated Immunosuppressive Properties by IL-10 and Regulatory T Cells Min Kyung Jung1, Shin Myung Kang2, Eui-Cheol Shin1* 1 Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, KAIST, Daejeon, 2Department of Internal Medicine, Gachon University Gil Hospital, Incheon, Korea Tel: 042-350-4286, E-mail: [email protected] Although lung diseases due to nontuberculous mycobacteria (NTM) are becoming prevalent worldwide, immunopathogenesis is poorly understood. To characterize immunological features of the patients with NTM lung diseases, we measured cytokines produced by PMA/ionomycin-stimulated peripheral blood mononuclear cells from NTM patients and NTM sensitin-reactive healthy controls by cytokine bead array. NTM patients had lower level of pro-inflammatory cytokines such as IFN-γ, TNF-α, and IL-17 than those of healthy controls. However, higher level of IL-10, a representative anti-inflammatory cytokine, was observed in NTM patients. These results suggest that NTM patients may be in state of suppressed inflammatory immune responses. To dissect regulatory mechanism of these immunosuppressed states of NTM patients, we analyzed regulatory T cell (Treg) frequency by flow cytometry. Frequency of + + + − Treg (CD4 CD25 Foxp3 CD127 ) was significantly increased in NTM patients compared with healthy controls. Additionally, expression of CD39 on Treg, activity marker of Treg, was markedly increased in NTM patients compared with healthy controls. This study suggests that suppressed inflammatory immune responses and high frequency of functionally activated Treg are immunological characteristics of patients with NTM lung diseases. Keywords: Nontuberculous mycobacteria (NTM), IL-10 and Regulatory T cell (Treg) Keywords: Acidic polysaccharide of Panax ginseng, Ionizing radiation, Radioprotection, Small intestines, Immune cells 76 The 2014 Fall Conference of the Korean Association of Immunologists Immune Response Regulation and Tolerance P-073 Neuro-Protective Effects of PLA2 by Suppression of Neuroinflammatory Responses in the 3xTg AD Mouse Model of Alzheimer’s Disease 1 1 2 1 Minsook Ye , Chanju Lee , Insop Shim , Hyunsu Bae * 1Department of Physiology, College of Korean Medicine, Kyung Hee University; Seoul, Acupuncture and Meridian Science Research Center, 2 College of Korean, Acupuncture and Meridian Science Research Center, College of Korean Medical Science Graduate School, Kyung Hee University, Seoul, Korea Tel: 02-961-0323, E-mail: [email protected] Alzheimer’s disease (AD) is a severe neurodegenerative disease for which there is currently no effective treatment. In the present study, we sought to determine whether PLA2 decreases the amyloid beta in the 3xTg mouse model of Alzheimer’s disease (AD). Treatment with PLA2 prevented increase of amyloid beta in the hippocampus. This neuro-protective effect of PLA2 was associated with microglial deactivation and reduction of CD4 T cell infiltration. Additionally, PLA2 treatment significantly increased the proportion of CD4+ CD25+ Foxp3+ Tregs in vivo. The increased proportion of Tregs by PLA2 treatment remained suppressive ex vivo. Therefore, our present studies suggest that modulation of peripheral immune tolerance by Treg may contribute to the neuroprotective effect of PLA2 in the 3xTg-AD mouse model of Alzheimer’s disease. This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korean government (Ministry of Education, Science, and Technology, MEST) (no. 2007-0054931). P-074 Poster Presentation NKT Cells Optimize Cytotoxic T Cell Activation by Direct Recognition of CD1d on T Cells Sejin Oh, Sojung Lim, Jung Hoon Shin, Se-Ho Park* Department of Life Sciences, Korea University, Seoul, Korea Tel: 02-3290-3646, E-mail: [email protected] Invariant NKT cells (iNKT cells) recognize foreign and endogenous glycolipid antigens that are presented by CD1d and essential for the optimal activation of cytotoxic T cells (CTLs). iNKT cell-dependent optimization of CTL activity is generally, but not conclusively, considered to be indirectly mediated by antigen-presenting cells (APC), which are stimulated by iNKT cells. In this study, we investigated the mechanism by which iNKT cells promote CTL responses. We found that CD8+ T cells begin to up-regulate the surface expression of CD1d upon antigen recognition. These CD1d-high CTLs became superior stimulators for iNKT cells when compared to CD1d-low CTLs. Activated iNKT cells, in turn, further stimulated CTLs dependent on the level of CD1d on the activated CTLs. This enhanced CTL activation by iNKT cells was even observed in APC-free conditions. Finally, we observed that CD1d-deficient CTLs were not able to reject target antigen-bearing tumor cells as well as CD1d+ CTLs. These results suggest that the activation of CTL is finely regulated by direct interaction of NKT cells with CD1d on activated CTLs. Keywords: Invariant NKT cell, Cytotoxic T cell, CD1d Keywords: Alzheimer’s disease (AD), 3xTg AD, PLA2, Microglia, Neuroinflammation P-075 ImT Exctrat Inhibits NLRP3 Inflammasome Activation and Anti-Inflammation Response in Raw 264.7 Macrophages Stimulated with Lipopolysaccharide Xiao Sun, Ji-Won Han, Do-Wan Shim, Na-Hyun Oh, Woo-Young Shin, Kang-Hyuck Heo, Tae-Bong Kang, Kwang-Ho Lee* Department of Biotechnology, College of Biomedical & Health Science, Research Institute of Inflammatory Diseases (RID), Konkuk University, Chungju, Korea Tel: 043-840-3613, E-mail: [email protected] ImT is an annual herbaceous plant that grows naturally along waterfronts in the mountains in almost every region of far easrt Asia. It has been used for detoxication and treatment of carbuncles and contusions in Chinese medicine. However, there has been no report about its anti-inflammasome and anti-inflammatory effects. In this study, we elucidated the effects of ImT on activation of inflammasomes and inflammtion. Inflammasome complexes are one of the central components of these processes through their regulation of IL-1βand IL-18. ATP, Nigericin and MSU-stimulated BMDM cells were used to study the regulatory effect of ImP on inflammasome activation. ImT attemuated interleukin (IL)-1βmaturation, caspase-1 activity, and NLRP3 gene expression in murine macophages. Pre-treatment with ImP inhibited the LPS-stimulated nitric oxide (NO) release, interleukin (IL)-6 production in Raw 264.7 cells. ImP also inhibited the LPS-stimulated STAT1 and IRF3 phosphorylation. In vivo animal sepsis model was employed to confirm the inhibitory effect of ImP on inflammatory reaction in vivo. ImP exhibited potent anti-inflammasome and anti-inflammation effect and may provide a valuable therapeutic strategy in treating various inflammation-mediated diseases. P-076 Weissella cibaria JW-15 Enhances Natural Killer cell Activity in vitro Kyung-Won Kang, Young-Joo Yi*, Sang-Myeong Lee* Division of Biotechnology, Chonbuk National University, Iksan, Korea Tel: 063-850-0843, E-mail: [email protected] Probiotics provide benefits in increasing host immune responses and protecting against infection. Recent studies reported that heat-killed probiotics showed significant immunomodulatory effects on the production of TNF-αand IFNγ. Natural killer (NK) cells play important roles in innate immune responses and inducing the spontaneous killing of tumor and virus-infected cells. However, the effects of heat-killed probiotics have not been studied on NK cells. Therefore, we evaluated the effect of heat-killed Weissella cibaria JW-15 (JW-15) isolated from Kimchi on NK cell activity and investigated the mechanisms underlying. In order to find out whether heat-killed JW-15 could increase NK cell proliferation and activation, we detected the proportion of NK cells by staining human peripheral blood mononuclear cells (hPBMC) for cell surface markers, followed by flow cytometry analysis. We conducted the cytotoxicity assay on NK cell. In addition, we performed intracellular cytokine staining to assess the secretion of the IFNγ, perforin, and granzyme B in NK cells. The cytotoxicity of NK cells was remarkably increased after treating hPBMC with JW-15. Also, JW-15 induced the secretion of IFNγin NK cells. The intracellular granule expression of perforin and granzyme B didn’t show significant differences in NK cells with or without JW-15. However, increasing mRNA expressions were confirmed in perforin, granzyme A & B and granulysin. Collectively, these data indicated that heat killed JW-15 enhances NK cell activity and can be used for immune-enhancement applications. Keywords: Probiotics, JW-15, Immunomodulatory effect, NK cells, IFNγ Keywords: Inflammasome, NLRP3, Inflammation The 2014 Fall Conference of the Korean Association of Immunologists 77 Poster Presentation P-077 Immune Response Regulation and Tolerance Establishment of Breast Cancer Mouse Models with TH1 and TH2 Types for the Study of Cancer Immunology P-078 Red Beet Protects Radiation-Induced Damage in Hematopoietic Stem/Progenitor Cells in Mice Ki Yeon Kim, Su Gang Kim, Sangjun Davie Jeon, Moon Gyo Kim* Areum Kim1, So Jin Bing1, Jinhee Cho1, Ginnae Ahn2, Youngheun Jee1* Department of Biological Sciences, Inha University Tel: 032-860-7728, E-mail: [email protected] 1 In order to study the immune responses during tumor progression and metastatic process, we established two mouse breast cancer models using immunocompetent C57BL/6 and Balb/c mouse strains. The syngenic breast cancer cell lines used for C57BL/6 and Balb/c are EO771 and 4T1, respectively. By using a simple metastatic assay through tail vein injection, both cell lines showed strong metastasis to the lungs of their hosts. These cells were also orthotopically implanted into mammary fat pad and followed for primary tumor growth and metastasis as well as for the survival and necrosis. The cell lines consistently express PRSS14/Epithin that plays critical roles in cancer metastasis. When the cells lose their expression of PRSS14/Epithin by introducing shRNA, the growth rates of the primary tumor were much slower and the metastasis was diminished in both TH1 and TH2 type mouse strains. In addition, the initiation of the tumor necrosis appears slower and the survival rates were increased. When KLH conjugated antigenic peptides derived from PRSS14/Epithin were immunized with various adjuvants, the antigen specific antibodies were efficiently produced. The degrees in reduction of metastasis correlated to the level of TH2 type antibody responses in both C57BL/6 and Balb/c models, indicating TH2 response is responsible for reducing metastatic process. Alum and MF59 adjuvants revealed high TH2 type while CFA adjuvants produced TH1 type antibody responses in both mouse strains. These two different types of mouse models with their coupled breast cancer cell lines and the modification techniques for immune responses are useful for the study of mechanism of immune modulation against both cancer progression and metastasis. Keywords: PRSS14/Epithin/matriptase, Immune response, Breast cancer model, Lung metastasis P-079 Ursolic acid Ameliorates Autoimmune Arthritis Via Suppression of Th17 and B cell Differentiation SeungYe Baek1, Jaeseon Lee1, Dong-Gun Lee1, Mi-Kyung 1 1,2 1,2 1 Park , Jennifer Lee , Seung-Ki Kwok , Mi-La Cho , 1,2 Sung-Hwan Park * 1 The Rheumatism Research Center, Catholic Research Institute of Medical Science, 2Divison of Rheumatology, Department of Internal Medicine, The Catholic University of Korea, Seoul, Korea Tel: 02-2258-6011, E-mail: [email protected] Objectives: Based upon the anti-inflammatory and anti-oxidative properties of Ursolic acid, we sought to examine the effects of UA in a mouse model of collagen-induced arthritis, and to identify the mechanisms underlying these effects. Methods: UA (150 mg/kg) was injected intraperitoneally three times a week beginning 14 days after the induction of CIA. The expression of proinflammatory cytokines and genes associated with oxidative stress in joints was measured by immunohistochemistry. The numbers of CD4+IL-17+, CD4+CD25+Foxp3+, and phosphorylated (p)STAT3+ cells were determined by confocal immunostaining, or by flowcytometric analyses. Serum antibody and mRNA expression were analysed by ELISAs and qRT-PCR, respectively. Results: UA reduced the incidence of clinical arthritis and joint inflammation significantly in mice with CIA. This clinical improvement was associated with a decrease in pSTAT3, along with reduced expression of IL-17 and RORγt, consistent with the decrease seen in Th17 cells, while the number of Treg cells was increased following UA treatment. There was also a marked reduction in CII-specific IgG production. Furthermore, expressions of the mRNA for Bcl-6, Blimp-1 and AID were markedly reduced in CD19+ B cells stimulated with IL-21 or LPS in the presence of UA. Conclusions: Our findings suggest that UA inhibits Th17 differentiation and promotes Treg cell proliferation by inhibiting the expression of RORγt and pSTAT3. UA also inhibits plasma cell differentiation, leading to a reduction in serum antibody levels. By targeting pathogenic Th17 cells and autoantibody production, UA may be useful for the treatment of autoimmune arthritis and other Th17-related diseases. College of Veterinary Medicine and Institute of Nuclear Science and Technology, Jeju National University, Jeju, 2Department of Marine Bio-Food Sciences, Chonnam National University, Yeosu, Korea Tel: 064-754-3374, E-mail: [email protected] Red beet (Beta vulgaris L.) has been demonstrated to present interesting biological and pharmacological properties, including antioxidant and anti-inflammatory properties. Herein, we elucidate the protective effects of red beet on radiation-induced damage in hematopoietic stem/progenitor cells. To confirm bone marrow and hematopoiesis ability of red beet, we irradiated (7 Gy) to mice with or without red beet (10 mg/kg). At 10 days after irradiation, red beet not only enhanced cell proliferation which was inhibited by radiation as shown in control group but also decreased DNA damages of immune cells. We found that red beet enhanced the survival of bone marrow cells along with increasing their number in the S phase of cell cycle simultaneously. Also, it was found that red beet treated group showed conspicuous improving of differentiation of hematopoietic stem cells (HSCs) into burst-forming units-erythroid (BRU-E) compared to untreated control group along with increased production of IL-3 from bone marrow cells. Red beet treated group also revealed enhancement in the level of hematocrit and hemoglobin in addition to the number of red blood cell in peripheral blood compared to control group. The results of this stydy suggest that red beet has the potential of maintaining bone marrow integrity and stimulating the differentiation of HSCs against irradiation in mice. This work (Grants No. C0146890) was supported by Business for Cooperative R&D between Industry, Academy, and Research Institute funded Korea Small and Medium Business Administration in 2013, and High Value-added Food Technology Development Program, Ministry of Agriculture, Fodd and Rural Affairs. Keywords: Red beet, Radiation, Hematopoietic stem cells P-080 ZYM-201 has Inhibitory Effects on LPS-stimulated B Cells in vivo and in vitro Ye Eun Lee, Soochan Kim, Woong-Jae Jung, Hyung Soo Lee, Mi-Yeon Kim* Department of Bioinformatics and Life Science, Soongsil University, Seoul, Korea Tel: 02-826-7192, E-mail: [email protected] ZYM-201 is a methyl ester of triterpenoid glycoside from Sanguisorba officinalis which has been used for treatment of inflammatory and metabolic diseases. In this study, immunomodulatory effects of ZYM-201 on B cells were examined in vitro and in vivo. When splenocytes were activated with lipopolysaccharide (LPS), the major population which had shown an increase in cell numbers was B cells. However, when the B cells were treated with ZYM-201 after LPS activation, their cell numbers and the expression of major costimulatory molecules, CD80 and CD86, were decreased. Furthermore, the effect of LPS, which induces activation of NF-κB, was abolished by ZYM-201: LPS-stimulated B cells showed decrease of phosphorylation after treatment of ZYM-201. The same results were shown in vivo experiments. These results suggest that ZYM-201 may play a role in the modulation of inflammatory responses through inhibiting NF-κB activation and downregulating the expression of costimulatory molecules on B cells. Keywords: ZYM-201, B Cells, inflammation, LPS Keywords: Ursolic acid, Rheumatoid arthritis, Th17 cells, Regulatory T cells, Osteoclastogenesis 78 The 2014 Fall Conference of the Korean Association of Immunologists Immune Response Regulation and Tolerance P-081 CIt Attenuates Inflammatory Response in Raw 264.7 Macrophages Stimulated with Lipopolysaccharide and Inhibits IL-1β Secretion in Bone Marrow Derived Macrophages through Inflammasome Regulation Young-Eun Ji, Ji-Won Han, Do-Wan Shim, Cheol-Hun Jang, Tae-Bong Kang, Kwang-Ho Lee* Department of Biotechnology, College of Biomedical & Health Science, Research Institute of Inflammatory Diseases (RID), Konkuk University, Chungju, Korea Tel: 043-840-3613, E-mail: [email protected] Chrysanthemum plant is summer perennial herb of the temperate forests of Korea and Japan. We have carried out the analysis of ethnopharmacological function of alcoholic extract of chrysanthemum plant (CIt), . Lipopolysaccharide (LPS)-stimulated Raw 264.7 murine macrophages were used to study the regulatory effects of CIt on inflammatory mediators such as nitric oxide (NO) and pro-inflammatory cytokines expression. Western blotting or ELISA techniques were employed to estimate protein levels. Pre-treatment with CIt inhibited the LPS-stimulated NO release, interleukin (IL)-6 in Raw 264.7 cells dose dependently. CIt also inhibited the LPS-stimulated iNOS and JNK phosphorylation. Innate immune responses have the ability to combat against infectious microbes and drive pathological inflammation. Inflammasome complexes are one of the central component of these processes through their regulation of IL-1βand IL-18. ATP, Nigericin and Silica-stimulated BMDM were used to study the regulatory effect of CIt on inflammasome activation. CIt attenuated IL-1βsecretion through attenuation of inflammasome activation induced by ATP, Nigericin and Silica in BMDM. Keywords: IL-1β, Inflammasome, Inflammation, LPS P-082 Poster Presentation Identification of Epitope Spreading of P. gingivalis HSP 60 into Human Autoantigens 1 2 1 Jeomil Choi *, Euikyong Jeong , Gil Sun Cha , Sung-Jo 1 2 Kim , Ho Sung Kang 1 Department of Periodontology, School of Dentistry, Pusan National 2 University, Dental Research Institute, Department of Molecular Biology, College of Natural Sciences, Pusan National University, Tel: 055-360-5200, E-mail: [email protected] Background: Intramolecular or intermolecular epitope spreading is one of the four mechanisms on how antigens of infectious organisms may propagate into various human autoantigens to trigger adaptive autoimmune responses. Pep19 is one of immunodominant epitope of P. gingivalis heat shock protein 60 that has demonstrated proatherogenic property by elaborating TH1 responses. However, defined mechanism on how immunization of Pep19 may mobilize helper T cells and elicit B-cell responses to multiple human HSP autoantigens. Aims: The aim of the present study was to identify and characterize the TH cell responses to multiple autoantigens in young adults and experimental animal model. Materials and Methods: Pep19 was immunized in wild C57BL/6 mouse for identification of T-cell and B-cell responses 4- and 8-weeks following the last boosting immunization. In healthy teens and early twenties, serum reactivity to multiple human HSP autoantigens as well as ox-LDL has been characterized as it related to reactivity to Pep19 of P. gingivalis HSP 60. Results: TH1 cell and B-cell reactivity to Pep19 has been closely associated to Pep19, Pep9, Pep14 of human HSP60 and to ox-LDL in young adults and experimental animal. Conclusion: Intramolecular and intermolecular epitope spreading of Pep19 into human autoantigens could be demonstrated implicating its proatherogenic mechanism on how periodontal infection may trigger autoimmune atherosclerosis. This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (NRF-2011-0015501) and 2014 Clinical Research Grant, Pusan National University Dental Hospital. Keywords: Epitope spreading, Atherosclerosis, Periodontitis, Autoimmunity, Porphyromonas gingivalis, Heat shock protein, Autoantigens P-083 Lactoferrin is as Potent as TGF-β1 in the Induction of Regulatory T Cells Sun-Jin Kim1, Goo-Young Seo1, Jeong-Min Lee1, Bo-Eun Kwon2, Hyun-Jeong Ko2, Woan-Sub Kim3, Pyeung-Hyeun 1 Kim * 1 Department of Molcular Bioscience, College of Biomedical Science, 2 Kangwon National University, ChunCheon, Laboratory of Microbiology and Immunology, College of Pharmacy, Kangwon National University, 3 ChunCheon, Korea, Department of Animal Life and Environmental Science, Hankyong National University, Anseong, Korea Tel: 033-250-8546, E-mail: [email protected] Lactoferrin (LF), a pleiotropic ion-binding glycoprotein, is known to modulate the immune response. Since we recently observed that LF acted like TGF-βin IgA B cell differentiation, we investigated whether LF affects regulatory T cell (Treg) differentiation. LF substantially enhanced Foxp3 transcription but not canonical transcriptional factors/cytokines of other T cell subsets (T-bet↓, GATA3↓, IL-17↓, IFN-β↓, and IL-4↓). + + We found that LF increased differentiation of CD4 Foxp3 Treg from naïve CD4+ T cells in dose-dependent manner. Further, LF-induced Treg suppressed the proliferation of CD4+T cells as much as TGFβ-induced Treg. Finally, in mouse DSS-induced colitis model, administration of LF ameliorated colitis symptoms and increased frequency of intestinal CD4+Foxp3+ Treg cells. Taken together, these results suggest that LF is a potent inducer of regulatory T cells. Keywords: Lactoferrin, Regulatory T cells, Immune tolerance, Colitis P-084 Murine Mesenchymal Stem Cell-Derived Exosomes Suppress Th17, but Not Th1 Differentiation Sun-Ho Lee1,2,3,4, Yong-Hee Kim1,3,4, Jun-Seop Shin1,3,4, 1,3,4 1,3,4 Byoung-Hoon Min , Jong-Min Kim , Sung jun 1,2,3,4 1,2,3,4 , Chung-Gyu Park * Gang 1 Department of Microbiology and Immunology, 2Department of Biomedical Sciences, 3Cancer Research Institute, 4Xenotransplantation Research Center, Seoul National University College of Medicine, Seoul, Korea Tel: 02-740-8311, E-mail: [email protected] Mouse mesenchymal stem cells (MSCs) alleviate the inflammation and the disease severity in experimental autoimmune encephalitis (EAE). But, the underlying mechanisms are not fully understood. In the present study, the exosomes which were originated from MSCs were tested for their immunosuppressive effects. Exosomes exhibited potent suppressive effects on T cell proliferation both through the cell cycle arrest and apoptosis. Specifically, treatment with MSC-derived exosome significantly reduced the level of RORγt and IL-17A expression at Th17 differentiation condition. However, the expression level of T-bet and IFN-γ showed no difference between the exosome-treated and control group under Th1 differentiation condition. To prove the effect of the exosomes in vivo, EAE was induced in four cohorts of mice and each group was given with exosome, exosome-enriched, exosome-depleted by methyl-β- cyclodextrin (MβCD) culture supernatant or saline in the disease initiation phase. The development of EAE and secretion of IL-17A was significantly suppressed by exosome or exosome-enriched culture supernatant, but not exosome-depleted culture supernatant and saline. In conclusion, exosomes secreted from MSCs can suppress T cell activation and specifically abrogate Th17 T cell differentiation in vitro and in vivo. Keywords: MSC, Exosomes, Th17 The 2014 Fall Conference of the Korean Association of Immunologists 79 Poster Presentation P-085 Immune Response Regulation and Tolerance TLR7 Agonist, Imiquimod, Directly Inhibits IgG1 and IgE Production through Suppressing Their Germline Transcriptions in B Cells Hee-Kyung Yoon, Yung-Choon Yoo, Junglim Lee, Seok-Rae Park* Department of Microbiology, College of Medicine, Konyang University, Daejeon, Korea Tel: 042-600-6497, E-mail: [email protected] Toll-like receptor 7 (TLR7) recognizes viral ssRNA in certain cell types, such as dendritic cells and macrophages, and leads to IFN-αproduction. B cells also express TLR7 but the role of TLR7 in B cells was not fully understood. In this study, we investigated the direct effect of in vitro stimulation with a selective TLR7 agonist, imiquimod (R837), on mouse B cell viability, antibody production, and isotype switching. Imiquimod enhanced cell viability by anti-CD40- and IL-4-stimulated B cells while it selectively diminished IL-4-induced IgG1 and IgE production in anti-CD40-activated B cells. We also found that imiquimod suppresses the expression not only of germline γ1 and εtranscripts (GLTγ1 and GLTε) but also of post-switch γ1 and εtranscripts (PSTγ1 and PSTε). Moreover, imiquimod decreased IL-4-induced GLγ1 and GLε promoter activities. Collectively, these results suggest that direct stimulation of B cell TLR7 suppresses IL-4-induced IgG1 and IgE class switching via inhibition of their GL transcriptions and thus result in reduced production of IgG1 and IgE. Keywords: TLR7, Imiquimod, B Cells, IgG1, IgE P-086 Modulation of Nuclear Receptor 4 A Activity in T Cells Ji-Hyeon Shin, Eun Sook Hwang* College of Pharmacy and Graduate School of Pharmaceutical Sciences, Ewha Womans University, Seoul, Korea Tel: 02-3277-4693, E-mail: [email protected] Three members of nuclear receptor (NR) subfamily 4 A, NR4A1 (Nur77, NGFI-B), NR4A2 (Nurr1), and NR4A3 (Nor1) belong to the steroid nuclear hormone receptor which binds to the gene promoter and enhancer in the nucleus. They are highly homologous in the DNA binding domain but very distinct in their activation domain. NR4A subsets are immediate-early genes induced by multiple stimuli and bind the consensus NGFI-B response elements sequence (NBREs) as monomers. Orphan receptors have shown to regulate various biological processes among which are cell survival and differentiation, apoptosis, inflammation and metabolism. Recent studies reported that NR4A subsets play a critical role in the regulation of development of regulatory T (Treg) cells through the regulation of FoxP3. Despite the importance of NR4A in immune system, it is largely unknown how the NR4A activity is controlled in T cell immune system. In this study, we have identified a novel NR4A modulator, TAZ (transcriptional co-activator with PDZ-binding motif) that plays a key role for the control of cell differentiation and organ development via interaction with different transcription factors through the protein-protein interaction domains such as WW and coiled-coil domains. We demonstrated that TAZ interacted with NR4As and modulated NR4A-mediated gene transcription. Interestingly, the NR4A1 and NR4A2-induced promoter activity were reciprocally regulated by the expression of TAZ. While NR4A2-mediated reporter activity was significantly increased by TAZ, the reporter activity driven by NR4A1 was suppressed by the expression of TAZ in a dose-dependent manner. Since the induction of Foxp3 expression is regulated by NR4A1 and NR4A2, Foxp3 expression in T cells could be modulated by TAZ. In addition, TAZ deficiency enhanced inflammatory T cell generation. These results suggest that TAZ may have an important role at the modulation of NR4A activity in T cells. The detailed molecular mechanisms and in vivo physiological meaning of NR4A and TAZ interaction remain to be characterized. Keywords: Nuclear receptor, NR4A, Treg, Foxp3, TAZ P-087 Immune Suppressive Effect of Cancer Cell-Associated VSIG4 on NKT Cytokine Production Eungchang Lee, Keunok Jung, Inhak Choi* Advanced Reseach Center for Multiple Myeloma, Department of Microbiology and Immunology, Inje University College of Medicine, Busan Inje University College of Medicine, Busan, Korea Tel: 051-890-6734, E-mail: [email protected] VSIG4 is a newly identified co-signaling molecule inhibits the T cell cytokine through complement receptor or a putative receptor on T cell. In previous study, we demonstrated that VSIG4 -expressing kupffer cells, a type of liver-resident macrophage, play a critical role in liver immune tolerance by inducing and maintaining liver T and NKT cell tolerance. Even though the expression of VSIG4 in plasmacytoma of mutitple myeloma patient was associated poor prognosis, the role of VSIG4 expressed on cancer cells was not elucidated yet. To determine the effect of tumor-associated VSIG4 on NKT cells, we generated VSIG4 epressing MOPC315.BM cell line (MOPC315.BM-VSIG4) using Lentivirus expression system. Injection of α-Galactosylceramide (GC)-loaded MOPC315.BM-VSIG4 cells into Balb/c mice significantly reduced IFN-g secretion of NKT cells compared to that of α-GC-loaded MOPC315.BM-MOCK cells. For in vitro assay, purified liver NKT cells of Balb/c mice were cocultured with MOPC315.BM-VSIG4 cells or MOPC315.BM-mock cells pulsed with α-GC. MOPC315.BM-VSIG4 cells suppressed NKT cell IFN-g production in vitro. Therefore, our results suggest that tumor cell-associated VSIG4 could serve a therapeutic target for cancer immunotherapy. Keywords: Immune checkpoint, VSIG4, NKT cell P-088 Immunosuppressive Effects of Gamma-Synuclein in Bone Marrow-Derived Dendritic Cells Seong-Mook Kang1, Mi-Hyoung Kim1, Kyung-Hee Song1, 1 1 2 Seung-Youn Jung , Jiyeon Ahn , Dae-Seog Lim , 1 Jie-Young Song * 1 Division of Radiation Cancer Research, Korea Institute of Radiological & Medical Sciences, Seoul, 2Department of Biotechnology, CHA University, Gyeonggi-do, Korea Tel: 02-970-1308, E-mail: [email protected] Dendritic cells (DCs) are the most potent antigen presenting cells as the initiator of cellular immune responses. Tumor cells produce several immunosuppressive factors to maintain their survival and then evade immune surveillance in tumor microenvironment. We previously identified significantly up-regulated several factors by irradiation on human breast cancer cells (MDA-MB-231 and MCF-7) using quantitative proteomic analysis. Among them, gamma-synuclein (SNCG, BCSG1), a member of synuclein protein family, is highly induced by irradiated breast cancer cells and already reported to be a marker for tumor progression. Here, we aim to investigate whether radiation-induced tumor-derived factors like SNCG affect immune responses surrounding tumors, mouse bone marrow-derived DCs were treated with recombinant SNCG protein in the absence or presence of LPS. SNCG altered the phenotypic and functional maturation of DCs by inhibiting the surface expression of co-stimulatory molecules, such as CD40 and CD86, and decreasing mRNA level of pro-inflammatory cytokines (IL-12, IL-23, IL-6 and IL-1β). To confirm the immunosuppressive effect of SNCG on DCs, SNCG-treated DCs were co-cultured with splenic T cells. SNCG-treated DCs marginally inhibited T cell proliferation as well as distinctively increased the production of TGF-βand the population of regulatory T cells (CD4+CD25+Foxp3+). These results demonstrated that SNCG released by irradiated tumors could play a role in immunosuppression through inhibition of DC maturation in tumor microenvironment. Keywords: Tumor microenvironment, Tumor-derived factor, Gamma-synuclein, Dendritic cells, Regulatory T cells 80 The 2014 Fall Conference of the Korean Association of Immunologists Immune Response Regulation and Tolerance P-089 Localization of Upstream DNA Regions Regulating T Cell Immunoglobulin and Mucin + Domain-3 Expression in CD4 T Cells P-090 Poster Presentation Down-Regulation of Tet2 Prevents TSDR Demethylation in IL2 Deficient Regulatory T Cells Su-jin Yun, Ka-jung Jun, Kuniharu Komori, Sun Park* Varun Sasidharan Nair, Kwon Ik Oh* Department of Microbiology, Ajou University School of Medicines, Suwon, Korea Tel: 031-219-5071, E-mail: [email protected] Department of Pathology, Hallym University College of Medicine, Chuncheon, Korea Tel: 033-248-2564, E-mail: [email protected] T cell immunoglobulin and mucin domain-3 (TIM-3) is expressed in vari+ ous immune cells and regulates immune responses of these cells. In CD4 T cells, TIM-3 expression is induced by activation signals especially in Th1 differentiation condition. In this study we localized regulatory regions in upstream DNA sequences of TIM-3 coding gene. We found the region of -144 bp ∼ -1 bp from transcription start site of TIM-3 as a minimal promoter. This region contains a putative AP-1 binding site and its mutation reduced luciferase reporter activity. When AP-1 is overexpressed, both TIM-3 proximal promoter activity and mRNA expression were increased. Among upstream 4 different CNS, increased dimethylation of histone H3 lysin4 (H3K4me2) indicating enhancer region was observed on CNS4 in Jurkat T cells and Th1 cells expressing TIM-3 but not in Th2 cells not expressing TIM-3. Furthermore, CNS4 increased TIM-3 minimal promoter activity only when two putative NF-κB binding sites in this region was not mutated. Taken together, our results suggest that minimal TIM-3 promoter region containing an AP-1 binding site and CNS4 containing NF-κB binding sites are important in TIM-3 transcriptional regulation. Stable expression of Foxp3 in regulatory T (Treg) cells is dependent on both intrinsic factors like epigenetic changes (demethylation) of Treg cell specific demethylation region (TSDR) and environmental cues like inflammations. Interleukin-2 (IL2) was reported to be one of the cytokines that give signals to Foxp3 stability but the underlying mechanism is still elusive. Here we show that IL2 and epigenetic changes in foxp3 locus are closely connected through tet methylcytosine dioxygenase 2 (Tet2) and, together help Treg cells to express Foxp3 stably. TSDR in foxp3 locus was not demethylated and Foxp3 expression was labile in IL2 deficient Treg cells, which was not restored by recombinant IL2, but correlated with the down-regulation of Tet2. Tet2 was up-regulated by TCR signaling and IL2 had a minimal effect. Rather, IL2 seemed to maintain the high level of Tet2 indirectly. Furthermore, over-expression of Tet2 restored TSDR demethylation in IL2 deficient Treg precursors. Collectively, our results suggest that up-regulation of Tet2 is required for Foxp3 stability and IL2 is required to maintain the high level of Tet2 during the thymic Treg development. + Keywords: TIM-3, Transcriptional regulation, CNS, CD4 T cell P-091 Upregulation of PD-1 on Regulatory T Cells Potentiates Their Suppressive Function during Chronic Viral Infection Hyo Jin Park1*, Joon Seok Park1, Yun Hee Jeong1, Chan 1 2 3 4 Hee Park , Byoung-Hee Lee , Lieping Chen , Jun Chang , 5 6 1 Doo Hyun Chung , Inhak Choi , Sang-Jun Ha * 1 Department of Biochemistry, College of Life Science & Biotechnology, Yonsei University, Seoul, 2Division of Biological Resources Coordination, National Institute of Biological Resources, Environmental Research Complex, Incheon, Korea, 3Department of Immunobiology, Yale University School of Medicine, New Haven, CT 06520, USA, 4Division of Life & Pharmaceutical Sciences, and Center for Cell Signaling & Drug Discovery Research, Ewha Womans University, Seoul, 5Department of Pathology, College of Medicine, Seoul National University, Seoul, 6Department of Microbiology and Immunology, Inje University College of Medicine, Busan, Korea Tel: 02-2123-7584, E-mail: [email protected] Regulatory T (Treg) cells act as terminators in the case of T cell immunity during the acute phase of viral infection; however, their roles in chronic viral infection are not completely understood. Here, we compared the phenotype and function of Treg cells during acute and chronic viral infection using lymphocytic choriomeningitis virus-infected mouse models. Chronic infection, unlike acute infection, led to induction of Treg cells and upregulation of programmed death-1 (PD-1). Treg cells isolated from chronically infected mice displayed greater suppressive capacity for inhibiting T cell proliferation and subsequent cytokine production than those from naïve or acutely infected mice. Suppression was contact-dependent and required PD-1 expression. We found that PD-1 signaling in Treg cells enhanced suppressive function by upregulating IL-10 and granzyme B. These findings establish PD-1 as a mediator of Treg cell suppressive function in the regulation of T cell responses and suggest a role of PD-1 expression on Treg cells, in addition to that on exhausted T cells, during chronic viral infection. Keywords: Regulatory T cells, Foxp3, Demethylation, Tolerance, Interleukin-2 P-092 Immune Suppression Function of Anti-Malarial Drugs through T Cell Inhibition Sera Oh, Yeon Ji Oh, Ji-Hyeon Shin, Eun Sook Hwang* College of Pharmacy and Graduate School of Pharmaceutical Sciences, Ewha Womans University, Seoul, Korea Tel: 02-3277-4693, E-mail: [email protected] Quinoline derivatives including chloroquine (CQ), amodiaquine (AQ), mefloquine, and primaquine have been widely used for preventing and treating malarial infection. CQ is the first major anti-malarial drug for treatment and infection of Plasmodium ovale and P. malariae and AQ, a similar drug to CQ is effective for preventing the infection of CQ-resistant P. vivax and P. falciparum. CQ and AQ have been identified to have potent anti-malarial and anti-inflammatory activity. However, the detailed molecular and cellular mechanisms are largely uncharacterized. In this study, we investigated whether anti-malarial drugs affect CD4+ T cell functions. The effect of CQ and AQ on the activation and proliferation of CD4+ T cells was assessed. CQ has no potent cytotoxic effect on CD4 T cell activation, whereas AQ displayed a strong cytotoxicity at the concentration over 10μM. Under 10μM concentrations, interestingly, AQ significantly suppressed T cell proliferation and increased the expression of p21 in a dose-dependent manner. Furthermore, AQ treatment resulted in decreased production of interferon-gamma (IFN-γ) while IL-2 production was not affected. Our results suggest that an antimalarial drug, AQ selectively inhibited T cell function though inhibition of cell proliferation and IFN-γproduction. Further studies on the characterization of the detailed anti-proliferative mechanisms of AQ and the in vivo functions of AQ in inflammatory disease are on-going. Keywords: AQ, CQ, CD4+ T cells, p21, IFN-γ Keywords: Programmed death-1, Regulatory T cells, LCMV The 2014 Fall Conference of the Korean Association of Immunologists 81 Poster Presentation P-093 Immune Response Regulation and Tolerance Novel Function of Irf8 in Suppression of Type I Immune Response by Regulatory T Cell Wonyong Lee, Gap-Ryol Lee* Department of Life Science, Sogang University Tel: 02-705-8458, E-mail: [email protected] P-094 Mitochondiral Cardiolipin Activates Dendritic Cells Enough to Activate Immune Responses 1,4 1,2,3,4 , Hye-Jung Moon1, Jung-Ah Cho , Tae-Joo Kim 1 1 5 Young-Joo Kim , Hye-Kyung Yoon , Polly Matzinger *, 1,2,3,4 Seung-Yong Seong * 1 Regulatory T cells (Treg cells) are small population of CD4+ T cells and take role in immunological suppression and tolerance. It has been suggested that Treg cells can be divided into multiple subsets which differs in the expression of distinct transcription factors and surface markers similar with effector T cells they target. We present transcription factor Irf8 as a transcription factor playing a role in a subset of Tregs. It is induced by Foxp3 and is important for the expression of chemokine receptor CXCR3, that is involved in cell migration and also commonly expressed in T helper 1 cells. Another transcription factor responsible for the expression of CXCR3, T-bet, was not affected by Irf8. Irf8-deficient mice showed significantly reduced CXCR3 expression in Treg cells and were impaired in Treg recruitment to the liver in αGal-C18-Cer-induced hepatitis model. Our results support that Irf8 in Treg cells controls Th1 immune response in T-bet independent mechanism. Keywords: Irf8, Treg Wide River Institute of Immunology, Seoul National University, Hongcheon, 2Department of Microbiology and Immunology, 3Department of Biomedical Sciences, 4BK21plus biomedical project, Seoul National University College of Medicine, Seoul, Korea, 5Laboratory of Cellular and Molecular Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA Tel: 033-3668-7651, E-mail: [email protected] Anti-phospholipid syndrome is one of autoimmune diseases with the high titers of anti-CL antibody. In this study, we investigated whether CL could activate dendritic cells (DCs) enough to induce the antigen-specific adaptive immune responses. Our results showed that CL could activate DCs via TLR2/CD14 receptors and signaling pathways as like as Gram(+) bacteria do. CL upregulated the expression of the co-stimulatory molecules on DCs in vivo and in vitro, and increased the production of several pro-inflammatory cytokines from DCs. The activated DCs were able to stimulate the antigen-specific activation and proliferation of T cells as well as antibody production of B cells in vivo. These findings show that CL could activate DC - T cell - B cell axis, suggesting a potential role of CL in contribution to pathogenesis of auto-immune diseases including anti-phospholipid syndrome. Keywords: Cardiolipin, Dendritic cells, Auto-immune disease, Antiphospholipid syndrome P-095 Age-Dependent Changes in Pattern Recognition Receptor and Cytokine mRNA Expression in Children with Otitis Media with Effusion Sang Hoon Kim1, Jae Yong Byun1, Moon Suh Park1, 1 2 Seung Geun Yeo , Dong Choon Park * 1 Department of Otorhinolaryngology-Head and Neck Surgery, School of 2 Medicine, Kyung Hee University, Seoul, Department of Obstetrics and Gynecology, St. Vincent's Hospital, College of Medicine, The Catholic University of Korea, Suwon, Korea Tel: 02-958-8474, E-mail: [email protected] Objective: To investigate age-dependent changes in expression of pattern recognition receptors (PRRs) and cytokines in pediatric OME. Material and Methods: Ninety five pediatric patients with OME were divided into 4 age groups: 0-2, 2-4, 4-7, and over 7 years. The presence of bacteria, and the levels of expression of mRNAs encoding toll-like receptor (TLRs), NOD like receptors (NLRs) and cytokines in middle ear fluid were assessed, as were their correlations with age, gender, presence of bacteria and accompanying disease. Results: Bacteria were detected in 32.6% of patients. The levels of expression of PRR and cytokine mRNAs tended to be lower in children aged 2-4 and 4-7 years. The levels of expression of TLR-2,-TLR-9, NOD-1, NOD-2, IL-1, IL-6, and TNF-α mRNAs in effusion fluid were significantly lower in these two groups than in children aged 0-2 and over 7 years (p<0.05 each). The levels of expression of TLR-4, TLR-5, TLR-9, and NOD-1 mRNAs were significantly lower in culture positive than in culture negative patients (p<0.05 each). However, the expression levels of PRR and cytokine mRNAs were unrelated to gender and accompanying disease (p>0.05 each). Conclusions: The levels of expression of PRR and cytokine mRNAs differed by age in children with OME. P-096 Organosulfur Inhibits NLRP3 Inflammasome Activation Huijeong Ahn, Jeeyoung Kim, Geun-Shik Lee* College of Veterinary Medicine, Kangwon National University, Chuncheon, Korea Tel: 033-250-8695, E-mail: [email protected] Methylsulfonylmethane (MSM) is an organosulfur compound and the health benefits associated with MSM include inflammation. Although MSM has been shown to have various physiological effects, no study has yet focused on inflammasome activation. The inflammasome is a multiprotein complex that serves as a platform for caspase 1-dependent proteolytic maturation and secretion of interleukin-1β(IL-1β). In this study, we tested the effect of MSM on inflammasome activation using mouse and human macrophages. In our results, MSM significantly attenuated NLRP3 inflammasome activation in lipopolysaccharide-primed macrophages, although it had no effect on NLCR4 or AIM2 inflammasome activation. Extracts of MSM-enriched vegetables presented the same inhibitory effect on NLRP3 inflammasome activation as MSM. MSM also attenuated the transcriptional expression of IL-1α, IL-1β, IL-6, and NLRP3. Taken together, these results show that MSM has anti-inflammatory characteristics, interrupts NLRP3 inflammasome activation, and inhibits pro-cytokine expression. We further confirmed the intracellular mechanism of MSM in relation to NLRP3 inflammasome activation, followed by comparison with that of DMSO. Both chemicals showed a synergic effect on anti-NLRP3 activation and attenuated production of mitochondrial reactive oxygen species. Thus, MSM is a selective inhibitor of NLRP3 inflammasome activation and can be developed as a supplement to control several metabolic disorders. Keywords: Methylsulfonylmethane, Organosulfur, Inflammasome, NLRP3, Interleukin-1beta Keywords: Otitis media with effusion, Toll-like receptor, Cytokine, Innate immunity 82 The 2014 Fall Conference of the Korean Association of Immunologists Innate Immune Response, Infection and Inflammation P-097 Polymer of the Amino Acid Glutamic Acid Inhibits Interleukin-1β Secretion via NLRP3 and AIM2 Inflammasome Activations Huijeong Ahn, Jeeyoung Kim, Geun-Shik Lee* College of Veterinary Medicine, Kangwon National University, Chuncheon, Korea Tel: 033-250-8695, E-mail: [email protected] Poly-gamma-glutamic acid (γ-PGA) is naturally produced from the bacteria Bacillus subtilis, and has been receiving increasing attention in the biomedical field due to its unique properties. The γ-PGA is unusual anionic polypeptide in which glutamate is polymerized via γ-amide linkages. Several studies have shown that γ-PGA induces a secretion of inflammatory cytokine, tumor necrosis factor-α, interukin-1β(IL-1β), and interferon-γ, and activated T cell, B cell, NK cell and immune cells. Based on the immune activity enhance effect of γ-PGA, we elucidated the effect of γ-PGA on inflammatory response, especially inflammasome activation. Inflammasome is a cytosolic sensory complexes, that controls processing and secretion of bioactive interleukin (IL)-1β, and is a central regulator of innate immune system and inflammation. In the present study, we elucidated that γ-PGA attenuated IL-1β secretion, readout of inflammasome activation, in the present of NLRP3, NLRC4 and AIM2 triggers. In addition, the extracts of γ-PGA-enrich Cheonggukjang were observed the similar anti-NLRP3, NLRC4 and AIM2 inflammasome properties like γ-PGA alone. Interestingly, γ-PGA up-regulated the transcriptional and translational expression of IL-1β implying that γPGA presented conflicting effects on innate immune system. Thus γPGA and Cheonggukjang could be suggested as a candidate to control inflammasome-mediated disorders such as type 2 diabetes. P-098 Poster Presentation The Inhibitory Effect of Paeoniflorin on Interleukin-1β Maturation Jeeyoung Kim, Huijeong Ahn, Geun-Shik Lee* College of Veterinary Medicine, Kangwon National University, Chuncheon, Korea Tel: 033-250-8683, E-mail: [email protected] Paeoniflorin, the major bioactive component of Paeonia lactiflora Pallas, has reportedly exhibited anti-inflammation, immune-regulation, anti-arthritis, anti-hepatitis, pain-relieving, neuromuscular blocking, and anti-hyperglycemia effects. Although immune-modulatory effects of PF have been progressively studied, the effect of paeoniflorin on inflammasome activation, newly defined proinflammaotry cytokine interleukin (IL)-1β maturation, has not been characterized yet. In this study, we elucidated the inhibitory effect of paeoniflorin on NLRP3, NLRC4 or AIM2 inflammasome activation human and murine macrophages. Paeoniflorin inhibited IL-1β secretions that were mediated by NLRP3, NLRC4, and AIM2 inflammasome activations and it also blocked the pyroptosis, caspase-1-mediated cell death, other indicator of inflammasome activation in BMDMs. Paeoniflorin further block the IL-1β producing mediated by the NLRP3 inflammasome activators, such as nigericin or alum in THP-1. We further confirmed the inhibitory mechanism of paeoniflorin on NLRP3 inflammasome. As results, paeoniflorin attenuated the rotenone- or KH7-induced IL-1βsecretion. Rotenone induces mitochondrial reactive oxygen species and KH7 inhibits adenylate cyclase. We demonstrated that Paeoniflorin has anti-inflammasome property with classical anti-inflammation characteristics. Keywords: Paeoniflorin, Inflammasome, NLRP3, NLRC4, Interleukin1beta Keywords: Poly-gamma-glutamic acid, Inflammasome, NLRP3, AIM2, Interleukin-1beta P-099 The Inhibitory Effect of Sulforaphane on NLRP3, NLRC4, and AIM2 Inflammasome Activations Jeeyoung Kim, Huijeong Ahn, Geun-Shik Lee* College of Veterinary Medicine, Kangwon National University, Chuncheon, Korea Tel: 033-250-8683, E-mail: [email protected] Sulforaphane is a natural isothiocyanate that is present in cruciferous vegetables such as broccoli and cabbage. Previous studies have shown that sulforaphane is effective in preventing carcinogenesis induced by carcinogens in rodents, which is related in part to its potent anti-inflammation properties. Although the effects of SFN on immunity have been progressively reported, its effects on inflammasome activation, multi-protein complex activates caspase-1 to produce the activated form of interleukin (IL)-1βand -18, have not been studied yet. In the study, we elucidated the inhibitory effect of sulforaphane on NLRP3, NLRC4 and AIM2 inflammasome activation using human and murine macrophages.We firstly checked the pro-inflammatory cytokines’ transcripts, such as IL-1α, IL-1β, IL-6, and TNFαin BMDMs after treated with sulforaphane. As expected, sulforaphane treatments significantly attenuated all of the cytokine expressions. In addition, sulforaphane significantly inhibited the ATP, a well-known NLRP3 inflammasome activator, mediating IL-1βsecretion, and ASC pyroptosome formation. Sulforaphane further block the IL-1β producing mediated by the other NLRP3 inflammasome activators, such as nigericin and alum. We further tested the effect of sulforaphane on other inflammasomes, NLRC4 and AIM2, using flagellin and dsDNA. Sulforaphane blocked the IL-1β secretion resulted from flagellin- and dsDNA- transfected macrophages. The extracts of sulforaphane-enriched cruciferous vegetables presented the same inhibitory effect on inflammasome activation like sulforaphane itself did. Finally, the underlying mechanisms for the anti-inflammasome activity of SFN were found to be a decrease in ATP-induced mitochondrial reactive oxygen species production. Taken together, sulforaphane may be one of the candidates to cure inflammasome mediating diseases. P-100 The Inhibitory Effect of β-D-glucan on NLRP3 and AIM2 Inflammasome Activations Jeeyoung Kim, Huijeong Ahn, Geun-Shik Lee* College of Veterinary Medicine, Kangwon National University, Chuncheon, Korea Tel: 033-250-8683, E-mail: [email protected] The inflammasome is a multiprotein complex in myeloid cells that serves as a platform for caspase-1 activation resulting in interleukin-1β(IL-1β) maturation. Polysaccharides from higher Basidiomycete mushrooms, mainly β-D-glucans, are considered to be potent bioactive compounds. β-D-glucans has been reported immune-modulatory properties however there is no reports on inflammasome activation. We elucidated the inhibitory effect of β-D-glucan on NLRP3, NLRC4 and AIM2 inflammasome activation using human and murine macrophages. As a result, the β-D-glucan significantly attenuated matured IL-1βsecretion resulted from NLRP3 or AIM2 inflammasime activations. However, βD-glucan did not effect on flagellin-induced IL-1βsecretion. In addition, mushroom extracts inhibited NLRP3 inflammasome however AIM2 and NLRC4 inflammasomes were differently regulated by each mushroom. Taken together, β-D-glucan was selective inhibitor for NLRP3 and AIM2 inflammasome activation. Keywords: Beta-D-glucan, Inflammasome, NLRP3, Interleukin-1beta Keywords: Sulforaphane, Inflammasome, NLRP3, Interleukin-1beta The 2014 Fall Conference of the Korean Association of Immunologists 83 Poster Presentation P-101 Innate Immune Response, Infection and Inflammation Obesity Reduces the Activation and Cytokine Production of Macrophage by Lipopolysaccharide Whajung Cho, Deuk-Ki Lee, Jae-Hwan Nam* Department of Biotechnology, The Catholic University of Korea, Bucheon, Korea Tel: 02-2164-4989, E-mail: [email protected] Obesity is a metabolic disease and is characterized by low-level chronic inflammation. Previous studies indicated that obese individuals were susceptible to virus infection and showed lower efficacy of vaccination than non obese people. In this study, we attempted to analyze the immunological environment of obesity using high fat diet (HFD) model. HFD increased body weight and epithermal fat mass. The proportion of activated B cell, T cell and macrophage was similar between HFD and regular fat diet (RFD) mouse. Th1 subpopulation was slightly increased in HFD mouse whereas the proportion of Th2 and Treg showed very little difference between both mice. In addition, proliferation and cytokine production of T cells when cells were stimulated with anti-CD3 and anti-CD28 antibodies in vitro did not differ in both mice. In the case of macrophages, expression levels of CD86 and MHC class II on freshly isolated peritoneal macrophage was similar but when cells were cultured in vitro, the proportion of CD86 expressed-macrophage was lower in HFD compared with RFD. Furthermore, LPS-induced IL-6 and TNF-α secretion was significantly reduced in macrophage isolated from HFD mouse. Taken together, our data suggested that insufficient functionality of macrophage in obesity may contribute to the reduction of protective immunity against pathogen. P-102 A Novel Natural Compound Alleviates Severe Sepsis 3 1 1,2,3 Youn-Hee Kim , Je-In Youn , Seung-Yong Seong * 1 Wide River Institute of Immunology, Seoul National University, Hongcheon, Department of Microbiology and Immunology, 3Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, Korea Tel: 02-740-8301, E-mail: [email protected] 2 The incidence of sepsis has steadily increased during the last few decades. However, the development of targeted therapy has been hampered because bacteria trigger the activation of multiple pro-inflammatory pathways. Here, we have investigated the effects of SNU304, a novel compound derived from natural products, on mouse models of sepsis. SNU304 has shown to protect mice against LPS induced sepsis and polymicrobial sepsis induced by CLP (cecal ligation and puncture). SNU304 administration also reduced blood pro-inflammatory cytokines level, such as TNF-α, IL-1β, IL-6, MCP-1, and IFN-γ. Blood pressure of the SNU304-treated mice showed a significant recovery from 4 hours after LPS challenge. SNU304 protects against acute kidney and liver damages in septic mice. SNU304 was also able to inhibit the expression of costimulatory molecules on DCs in LPS-induced sepsis mice, which might be involved with alleviation of the physiological outcome of sepsis. In conclusion, SNU304 mitigated the development of severe sepsis by modulating immune regulatory cells. Keywords: Sepsis, MDSC, Pro-inflammatory cytokine Keywords: Obesity, Macrophage, LPS, IL-6, TNF-alpha P-103 OxLDL Induces Endoglin Expression through TLR-dependent Pathway in Macrophages Young-Saeng Jang, In-Hong Choi* Department of Microbiology, Institute for Immunology and Immunological Diseases, Yonsei University College of Medicine, Seoul, Korea Tel: 02-2228-1821, E-mail: [email protected] Atherosclerosis is a chronic inflammatory disease of the arteries. Recent evidences have indicated that toll-like receptor (TLR)-dependent signaling plays key roles in development of atherosclerosis through the induction of cholesterol-laden foam cell formation and inflammatory response. However, target genes and endogenous ligands of TLRs that may be relevant to atherosclerosis are not delineated yet. In this study, we introduced endoglin (ENG), which is involved neoangiogenesis in advanced atherosclerotic lesions, as a target gene of TLR-signaling in atherosclerosis. Among various TLR agonists, TLR2 agonist enhanced the expression of membrane and soluble form of ENG on PMA-treated monocyte in a dose- and time-dependent manner. Interestingly, oxLDL also induced two forms of ENG expression, which was inhibited by blocking of TLR2 signaling. The experiments using inhibitors revealed that NF-κb and p38MAPK such as TLR-downstream signaling are involved in oxLDL-induced ENG expression. These results suggest that oxLDL induces ENG expression through TLR-dependent signaling. Consequently, these findings provide that ENG is an important insight into the novel target gene of TLR signaling in oxLDL-relative atherosclerosis. Keywords: Atherosclerosis, Endoglin, TLR, Oxidized LDL P-104 The Anti-Inflammatory Effects of Salicornia herbacea L. Ethanol Extract on Lipopolysaccharide-Stimulated RAW 264.7 Cells Mi-Rae Choi, Eun-Sil Ko, Ji-ye Lim, Jea-Ran Kang, Seung-Mi Hwang, Su-Beam Han, Kyung-Min Choi, Jeong-Dan Cha* Institute of Jinan Red Ginseng, Jinan, Korea Tel: 063-432-0944, E-mail: [email protected] Salicornia herbacea L. (S. herbacea) grows in salt marshes and salt fields along the seashores. Recent study on the effects of SH been known that diabetes has improved, antioxidant and anti-inflammatory. However, only a few studies have been carried out on anti-inflammatory effects. This study examined the inflammatory inhibition effects of S. herbacea ethanol extract (SHEE) on the lipopolysaccharide (LPS)-stimulated RAW264.7 cells. The SHEE significantly inhibited the production of NO in the LPS-stimulated RAW 264.7 cells, which was mediated by the down-regulation of inducible NO synthase (iNOS) expression but not by its direct cytotoxic activity. The SHEE was decreased as protein expression of Iκκα/β, nuclear translocation of p65, and subsequent activation of NF-κB as well as phosphorylation of p38 MAPKs at a dose-dependent manner. Moreover, MAPK inhibitor, p38 significantly reduced the production of NO on LPS-stimulated RAW 264.7 cells. In conclusion, we suggest that S. herbacea ethanol extract inhibits the expression and production of inflammatory mediators by blocking the MAPK-mediated pathways and inhibiting the activation of NF-κB. This paper was supported in part by research funds of Bio-Industry Technology Development Program (MFAFFRK 311013-03). Keywords: Salicornia herbacea L., Anti-inflammatory effects, Inflammatory mediators, NF-κB, MAPKs pathways 84 The 2014 Fall Conference of the Korean Association of Immunologists Innate Immune Response, Infection and Inflammation P-105 Orphan Nuclear Receptor Small Heterodimer Partner Is Translocated to Mitochondria in Macrophages after Inflammasome Stimulation Soo Yeon Kim1,2, Chul-Su Yang3, Eun-Kyeong Jo1,2* 1 Department of Microbiology, 2Infection Signaling Network Research Center, Chungnam National University School of Medicine, Daejeon, 3 Departmen of Molecular and Life Science, College of Science and Technology, Hanyang University, Ansan, Korea Tel: 042-580-8243, E-mail: [email protected] Small heterodimer partner (SHP; also known as NR0B2) is an orphan member of the nuclear receptor (NR) superfamily. The inflammasome is a large multimeric protein complex composed of NOD-like receptor proteins and adaptors that triggers caspase-1 activation, leading to maturation of the proinflammatory cytokines interleukin (IL)-1βand IL-18. Our previous study showed that SHP is localized in cytosols and essentially involved in negative regulation of toll-like receptor-induced signaling. However, little is known about the intracellular localization of SHP in macrophages after NLRP3 inflammasome stimulation. In this study, we examined the intracellular translocation of SHP in macrophages after NLRP3 inflammasome stimulation. We found that NLRP3 inflammasome stimulation led to translocation of SHP and NLRP3 to the mitochondrial fraction. Analysis of confocal microscopy data also supported mitochondrial translocation of SHP after NLRP3 inflammasome activation. In response to NLRP3 inflammasome stimulators, the SHP-NLRP3-ASC complex initially localized to the cytosolic fraction, before translocating to the mitochondrial fraction, in both primary macrophages and THP-1 cells. Triggering NLRP3 inflammasome stimulation induced a sequential interaction of NLRP3 with SHP and then, with ASC in the mitochondrial fraction. These results show that triggering NLRP3 inflammasome activation led to a translocation of SHP into mitochondria. Ongoing and future studies are being conducted in the functional analysis of SHP translocation into mitochondrial fraction in macrophages. P-106 Poster Presentation The Mycobacterium avium subsp. paratuberculosis MAP1305 Polarizes Dendritic Cell-Mediated T-Cell Proliferation Via Toll-Like Receptor 4 Kyung Tae Noh* Department of Infectious Diseases Research, Armed Forces Medical Research Institute, Daejeon, Korea Tel: 042-878-4799, E-mail: [email protected] In this study, we revealed that Mycobacterium avium subsp. paratuberculosis MAP1305 induces the maturation of bone marrow-derived dendritic cell (BMDC), a representative antigen presenting cell (APC). Also, MAP1305 provoked the production of pro-inflammatory cytokines (Interleukin (IL)-6), tumor necrosis factor (TNF)-α, IL-1β, and IL-12p70), and reduced the endocytic capacity of DC, which is the phenomena of mature DC. In addition, we found that MAP1305-mediated −/− DC maturation is via Toll like receptor 4 (TLR4), using TLR4 DC. It also activated MAPKs signaling pathway, which is essential for DC maturation Furthermore, MAP1305-treated DCs elicited naïve T cells to polar+ + ized CD4 and CD8 T cells, which would be crucial for Th1 polarization of the immune response. Taken together, M. avium subsp. paratuberculosis MAP1305 regulates innate immune responses through + + DC-mediated proliferation of CD4 and CD8 T cells. Keywords: Dendritic cells, MAPKs, MAP1305, Mycobacterium avium subsp. paratuberculosis, Th1 polarization Keywords: SHP, NLRP3, Inflammasome P-107 TLR2-Induced MMP9 Activation Compromise Blood Brain Barrier and Enhances Brain Damage in Collagenase-Induced Intracerebral Hemorrhage 1 1 1 Hyunjung Min , Jinpyo Hong , Yong Ho Jang , Kyung Il 1 2 1 Kwak , Soojin Lee , Sung Joong Lee * 1 Department of Neuroscience and Dental Research Institute, School of Dentistry, Seoul National University, Seoul, 2Department of Microbiology, School of Systems Biology, Chungnam National University, Daejeon, Korea Tel: 02-740-8649, E-mail: [email protected] P-108 Retnla Overexpression Attenuates Allergic Inflammation of the Airway Mi-Ran Lee1, Da Hee Shim2, Jihye Yoon2, Hyung Seok Jang2, Se-Woong Oh3, Suk Hyo Suh4, Goo Taeg Oh1, 2 Jae-Hoon Choi * 1 Department of Life Sciences, Ewha Womans University, Seoul, Korea, Department of Life Science, College of Natural Sciences, Research Institute for Natural Sciences, Hanyang University, Seoul, 3Yuhan Research Institute, Yuhan Corporation, Yongin, 4Department of Physiology Medical School, Ewha Womans University, Seoul, Korea Tel: 02-2220-2540, E-mail: [email protected] 2 Innate immune response plays an important role in the pathogenesis of cerebral ischemia/reperfusion injuries. Recent studies have shown that Toll-like receptor 4 (TLR4) is involved in the innate immune responses after cerebral ischemia/reperfusion injuries, yet, the role of TLR2 in stroke or the mechanisms of its function have not been resolved. In this study, we tested it in a collagenase-induced mouse intracerebral hemorrhage (ICH) model using TLR2 knock-out (KO) mice. To induce ICH, collagenase or blood was injected into the right caudate putamen in 8-10 week old male mice. TLR2 expression was upregulated in the ipsilateral hemorrhagic tissues of the collagenase-injected mice. Brain injury volume and neurological deficits following ICH were reduced in the TLR2 KO mice as compared to the wild type (WT) mice. Heterologous blood-transfer experiments show that TLR2 signaling in the brain-resident cells, but not leukocytes, contributes to the injury. In our study to elucidate underlying mechanisms, we found that damage in the blood-brain barrier (BBB) integrity following the ICH was attenuated in the TLR2 KO mice compared to the WT mice, which may be due to reduced MMP-9 activation in brain astrocyte. The reduced BBB damages accompanies with reduced neutrophil infiltration and proinflammatory gene expression the injured brain parenchyma, which may account for the attenuated brain damage in the TLR2 KO mice after ICH. Conclusively, these data demonstrate that TLR2 contributes to brain injury following ICH by compromising BBB through activating MMP-9 in brain. Resistin-like molecule alpha (Retnla), also known as ‘Found in inflammatory zone 1’, is a secreted protein that has been found in bronchoalveolar lavage (BAL) fluid of ovalbumin (OVA)-induced asthmatic mice and plays a role as a regulator of T helper (Th)2-driven inflammation. However, the role of Retnla in the progress of Th2-driven airway inflammation is not yet clear. To better understand the function of Retnla in Th2-driven airway inflammation, we generated Retnla-overexpressing (Retnla-Tg) mice. Retnla-Tg mice showed increased expression of Retnla protein in BAL fluid and airway epithelial cells. Retnla overexpression itself did not induce any alteration in lung histology or lung function compared to non-Tg controls. However, OVA-sensitized/challenged Retnla-Tg mice had decreased numbers of cells in BAL and inflammatory cells accumulating in the lung. They also showed a reduction in mucus production in the airway epithelium, concomitant with a decreased Muc5ac level. These results were accompanied by reduced levels of Th2 cytokines, including interleukin (IL)-4, IL-5, and IL-13, with no effect on levels of OVA-specific immunoglobulin isotypes. Furthermore, phosphorylation of ERK was markedly reduced in the lungs of OVA-challenged Retnla-Tg mice. Taken together, these results indicates that Retnla protects against Th2-mediated inflammation in an experimental mouse model of asthma, suggesting that therapeutic approaches to enhance the production of Retnla or Retnla-like molecules could be valuable for preventing allergic lung inflammation. Keywords: TLR2, Intracerebral hemorrhage, Astrocyte, MMP-9, Bloodbrain barrier Keywords: Retnla, Asthma, Inflammation, Macrophage The 2014 Fall Conference of the Korean Association of Immunologists 85 Poster Presentation P-109 Innate Immune Response, Infection and Inflammation Gingerol-Mediated Anti-Bacterial Immune Responses 1 2 2 Seong-A Ju , Tam Nguyen , Yea-Sol Lee , Jun-Hyeong 3 2 2 Bae , Thu-Ha T. Nguyen , Byung-Sam Kim * 1 Biomedical Research Center, Ulsan University Hospital, University of 2 Ulsan College of Medicine, Department of Biological Sciences, University of Ulsan, Ulsan, 3College of Life Sciences and Biotechnology, Korea University, Seoul, Korea Tel: 052-259-2350, E-mail: [email protected] P-110 Pochonin D Enabled Mice to Overcome Insufficient Protection Against Rhinovirus Infection Aeri Shim, Jae-Hyoung Song, eun-hye Hong, Bo-Ra Lee, Bo-Eun Kwon, Hyun-Jeong Ko* Laboratory of Microbiology and Immunology, College of Pharmacy, Kangwon National University, Chuncheon, Korea. Tel: 033-250-6923, E-mail: [email protected] Sepsis, a leading cause of death worldwide, involves concomitant expression of an overzealous inflammatory responses and inefficient bacterial clearance. 6-Gingerol, an aromatic polyphenol and the most pungent constituent of ginger (Zingiber officinale), has been found to possess many interesting pharmacological and physiological activities, such as anti-inflammatory, antioxidant, anti-tumor-promoting activities. We investigated the effects of 6-gingerol administration in septic mice subjected to polymicrobial sepsis. The model of polymicrobial sepsis was induced by cecal ligation and puncture (CLP) in BALB/c mice. 6-Gingerol treated mice developed less severe sepsis, lower mortality compared to control mice. Consistent with this data, 6-gingerol-treated mice had improved bacterial clearance in their organs, lower level of inflammatory cytokines, higher numbers of infiltrated neutrophils and macrophages in infected organs. In addition, 6-gingerol-treatment inhibited apoptosis of neutrophils and macrophages. Depletion of macrophages abrogated the effects of 6-gingerol in septic mice, indicating macrophages might be an effector cells producing protective effect of 6-gingerol in anti-bacterial immune responses. Our results suggest that 6-gingerol can be used in the treatment of clinical sepsis to prevent the detrimental effects of this disease. Human rhinoviruses (HRVs) is a positive single-stranded RNA virus belongs to the Piconaviridae family. They are divided into three distinct species: type A, type B, and type C and over 100 immunologically non-crossreactive HRVs serotypes. HRVs infection in human usually causes common cold and acute infectious illness, but sometimes they are associated with asthma exacerbations and viral upper respiratory infection. Despites of the highest in incidence and occasional complications of chronic bronchitis and exacerbations of reactive airway disease, there was no approved medication for the treatment of rhinovirus infection. HSP90, a 90kDa heat shock protein, is a highly abundant, essential, and evolutionary conserved molecular chaperones. There are two types of cytoplasmic isoforms of Hsp90 in mammals. Recent studies have demonstrated that Hsp90 inhibitors had broadest-spectrum antiviral activity. Several Hsp90 inhibitors have been studied for the development antiviral activity in vitro many viruses, but there are no studies for HRVs. In this study, we assessed the anti-HRV activity of Pochonin D, a well-known Hsp90 inhibitor, in vitro and in vivo and found that it has significant antiviral effect against HRV1B from reduced inflammation cytokines, virus titer, Histology, inflammatory immune cell analyzed. In addition, we found that Pochonin D affect virus replication and inhibit the HRV1B infection-associated lung inflammation. Keywords: 6-Gingerol, Polymicrobial sepsis, Anti-bacterial immune responses Keywords: Rhino Virus, Pochonin D, Antiviral P-111 Populational Dissection of T Helper (Th) Subsets in the Intestines of Mouse Infected with Vibrio vulnificus P-112 2B4 Receptor Is Essential to Host Defense Against L. pneumophila Pulmonary Infection Arim Lee, Tae Sung Kim* Bonggoo Park, Jung Eun Lee, Ga Young Park, Ji Young Kim, Kwang Hee Kim, Kyung-Mi Lee* Division of Life Sciences, College of Life Sciences and Biotechnology, Korea University, Seoul, Korea Tel: 02-3290-3416, E-mail: [email protected] Global Research Lab, Department of Biochemistry and Molecular Biology, Korea University College of Medicine, Seoul, Korea Tel: 02-920-6253, E-mail: [email protected] Vibrio vulnificus is a gram-negative bacterium, which causes fatal septicemia, gastroenteritis as a result of consumption of contaminated seafood or wound infection. It is known that V. vulnificus penetrates into the intestinal epithelial barrier and stimulates an inflammatory response in the adjacent mucosa. However, the exact effect of V. vulnificus infection on the intestinal immune system is not well understood. In the present study, we investigated the susceptibility of mice to oral infection with V. vulnificus and the course of infection as well as the associated Th subset responses in the small intestine. C57BL/6 mice were intragastrically inoculated with various colony-forming unit (CFU) of V. vulnificus. After 4 days of the infection, lymphocytes were isolated from spleen, mesenteric lymph nodes (MLN), peyer’s patch (PP), and lamina propria (LP) in V. vulnificus-infected and uninfected mice. Populations of Th subsets were determined by staining with specific mAbs against IL-4, IL-17, IFN-γ, and Foxp3, followed by flow cytometric analysis. Th17 cell population was higher in MLN, PP cells of mice infected with 107 CFU of V. vulnificus compared to that in uninfected mice. Th1 subset population was high7 er in LP, whereas it remained unchanged in PP after infection with 10 CFU of V. vulnificus. In mice infected with 108 CFU of V. vulnificus, Th17 subset population was increased in MLN, PP and LP, and Th1 subset population was increased in MLN, PP, and LP. These results suggest that understanding of intestinal Th cell-mediated immune response in mouse models of oral infection with V. vulnificus will be helpful to develop prophylactic and therapeutic agents against diseases caused by V. vulnificus. 2B4 receptor, a SLAM family member, is essential to a variety of immune responses against tumors and viral infections, so we’d like to investigate roles of 2B4 receptor against legionella infections. Legionella pneumophila is Gram-negative intracellular pathogen causing Legionnaires’ disease in humans. The pathogen multiplies in various host cells such as macrophages, dendritic cells and alveolar epithelial cells. Given that NK cells, CD4+ and CD8+ T cells are essential to innate immune defense against legionella infection as well as macrophages and neutrophils, 2B4 receptor must be involved in clearance of legionella from lungs during Legionellosis, since immune cells often express 2B4 receptor. Interestingly, in our pulmonary infection model using L. pneumophila, 2b4-/- mice have shown higher bacterial loads and inflammatory responses such as more neutrophils and macrophages compared to control. Thus, depletion out of and adoptive transfer into 2b4-/- mice would help us to identify which immune cells play major roles in defeating Legionellosis. BAL analysis showed slightly higher levels of IL-6, IFN-γ and TNF-α in 2b4-/- mice than controls, which means significant cytokine inductions happened before day 2. On day 2, cytokine levels such as IFN-γ, IL-6, MCP-1 and TNF-α appeared higher in blood serum of 2b4-/- mice than control ones, although their levels are relatively low indicating that their production also culminated in earlier times. In histology analysis, 2b4-/- mice had more severe pathologies in their lungs than controls, which are more hyaline membranes in blood vessels, more interstitial infiltration of inflammatory cells and necrotic abscess. Currently, there is no specific antibody to activate 2B4 receptor, thereby immune boosting host defense against L. pneumophila. Therefore, screening 2B4 activating antibodies or chemical agents would lead to novel therapeutics to control Legionella infections especially in immunocomprised patients. Keywords: V. vulnificus, Th subset, mouse, in vivo, intestine Keywords: 2B4 receptor, Legionella, NK cell 86 The 2014 Fall Conference of the Korean Association of Immunologists Innate Immune Response, Infection and Inflammation P-113 Withaferin A Reduces Inflammatory Responses Induced by Helicobacter pylori and Induces Apoptosis in Gastric Epithelial Cells Green Kim1, Tae-Hyoun Kim2, Min-Jung Kang1,3, Jin-A Choi1, Da-Young Paik1, Ik-Rae Lee1, Min-Gyu Kim1, Sang-Seop Han4, Bo-Yeon Kim3, Sang-Muk Oh1, 1 1 5 Kyung-Bok Lee , Dong-Jae Kim , Jong-Hwan Park * P-114 Poster Presentation Recruited M2 Macrophage Contribute to the Differentiation of Fibroblast and the Transition of Epithelial Cell in Radiation-Induced Pulmonary Fibrosis Hae-Ran Park*, Sung-Kee Jo, Uhee Jung 1 Research Division for Biotechnology, Jeongeup Campus of Korea Atomic Energy Research Institute (KAERI), Jeongeup, Korea Tel: 063-570-3222, E-mail: [email protected] Withaferin A, a withanolide purified from Withania somnifera, has been known to exert anti-inflammation effect and anti-tumor activity. But such inhibitory effects of withaferin A on gastric diseases have not been evaluated. Gastric cancer is one of the leading causes of death in the world and Helicobacter pylori-mediated inflammatory response is closely related to promotion of gastric cancer. In this study, we determined the effect of withaferin A on inflammation induced by H. pylori infection and proliferation of AGS cells, a gastric cancer cell line. Sub-cytotoxic level of withaferin A reduced IL-8 production in AGS cells in response to H. pylori. As well, withaferin A inhibited the activation of NF-κB, but not MAPKs in the cells infected with H. pylori. Such inhibitory effect is not associated with bactericidal property of withaferin A. Moreover, in high concentrations, withaferin A inhibited the growth of AGS cells. In flowcytometry analysis using Annexin V/PI staining, withaferin A led to the increase of apoptotic cells population. In addition, expression of cleaved PARP, caspases-3, -7, and -9 were significantly increased and bcl-2, an anti-apoptotic protein, was decreased by withaferin A in a dose-dependent manner. In conclusion, withaferin A inhibited H. pylori-induced production of IL-8 at low concentration and growth of gastric cancer cells by inducing apoptosis at high concentration. These findings suggest that withaferin A can be a potential therapeutic agent for gastric inflammation and cancer. Radiation-induced pneumonitis and pulmonary fibrosis are common dose-limiting complications in patients receiving radiotherapy for lung, breast, and lymphoid cancers. Fibrosis is defined as an excessive deposition of extracellular matrix (ECM) components, such as collagen, as a consequence of proliferation and activation of myofibroblasts. Recently, myofibroblasts were believed to derive from resident fibroblasts, circulating fibrocytes (derived from bone marrow stem cells), or injured epithelial cells. In our previous study, thoracic irradiation leads to accumulate macrophage into the lung. In this study, we investigated the characteristics of macrophage infiltrated after irradiation, and the role of M2-polarized macrophage in the differentiation of fibroblast and the transition of epithelial cell. After anesthesia, the whole thorax of C57BL/6 mice was irradiated at 14 Gy. At 4 and 6 months after irradiation, the CCL-2, CCL-3, and CXCL-10 were upregulated in the lung of irradiated mice, which leads to high attraction of macrophages into the lung following irradiation. In thoracic irradiated mice, macrophages infiltrated into the lung expressed the high levels of Mac-3 antigens and upregulated the hallmark of alternatively activated M2 macrophages such as arginase (Arg)-1 and macrophage mannose receptor (CD206). M2 macrophages induced the MLE12 to undergo phenotypic conversion to form fibroblast-like cells, called the epithelial-to-mesenchymal transition (EMT), which leads to the upregulation of EMT markers such as α-SMA, fibronectin, N-cadherin, vimentin, snail, and slug. Also, M2 macrophages induced the high level expression of collagen I in NIH-3T3. M2-polarized macrophages secreted the high level of TGF-β and IL-10 known as inducer of EMT. Although the actions of M2 macrophages may not account for the entirety of radiation-induced pulmonary fibrosis, we though that recruited-M2 macrophages play an important regulatory role during this process. Keywords: Helicobacter pylori, AGS cell, Withaferin A, Inflammation, Apoptosis Keywords: Radiation, Pulmonary fibrosis, M2 macrophage, Myofibroblast, Epitehlial-to-mesenchymal transition Department of Biochemistry, College of Medicine, Konyang University, 2 Daejeon, Laboratory Animal Medicine, College of Veterinary Medicine, Seoul University, Seoul, 3World Class Institute, Korea Research Institute of Bioscience and Biotechnology, Cheongwon, 4Graduate School of Pre5 Clinical Laboratory Science, Konyang University, Daejeon, Department of Laboratory Animal Medicine, College of Veterinary Medicine, Chonnam National University, Gwangju, Korea Tel: 062-530-2834, E-mail: [email protected] P-115 Involvement of Mast Cells in Inflammation by Trichomonas vaginalis via Crosstalk with Benign Prostatic Hyperplasia Epithelial Cells Jung-Hyun Kim1, Ik-Hwan Han1, Hyun-Ouk Song2, 3 1 1 Yong-Hoon Chung , Myoung-Hee Ahn , Jae-Sook Ryu * 1 Department of Environmental Biology and Medical Parasitology, Hanyang 2 University College of Medicine, Seoul, Department of Parasitology, School of Medicine, Catholic University of Daegu, Daegu, 3Department of Microbiology, Hanyang University College of Medicine, Seoul, Korea Tel: 02-2220-0683, E-mail: [email protected] It is known that chronic inflammation may have a role in pathogenesis of benign prostatic hyperplasia and prostate cancer. Mast cells have been detected in chronic inflammatory infiltrate of the prostate. It therefore seemed possible that the prostate epithelial cell-trichomonad reaction might affect the activity of mast cells present in the lamina propria of the prostate mucosa. In this study, we tested whether the culture supernatant of benign prostatic hyperplasia epithelial cell (BPH cell) incubated with Trichomonas vaginalis (BTCM) could stimulate mast cells. When BPH cells were incubated with live trichomonads, CCL2, CXCL8 and IL-1β productions increased in the BTCM. When the BTCM was added to mast cell (HMC-1 cell), β-hexosaminidase and CXCL8 productions increased. The BTCM also induced increased migrations of mast cell and THP monocyte. Moreover, the culture supernatant of HMC-1 cells incubated with BTCM (M-BTCM) showed increased chemotactic activity for mast cell and monocyte compared with that of BTCM. Taken together, the inflammatory mediators made by BPH epithelial cells in response to activation by T. vaginalis activate and attract mast cells. Our results suggest that BPH epithelial cells play a role in the infiltration of mast cells in T. vaginalis infection. Keywords: Trichomonas vaginalis, Benign prostatic hyperplasia epithelial cells, Mast cell, Inflammation P-116 MyD88-Dependent Protective Mechanism of Alveolar Macrophages in Mice Infected with Respiratory Syncytial Virus A2 Ji Eun Hong1, Sung-Moo Park1,2, In Su Cheon1,3, Yoon 1 4 3 5 Chul Kye , Chang Jun , Man Ki Song , Seung-Hyun Han , 1,2 Cheol-Heui Yun * 1 Department of Agricultural Biotechnology and Research Institute for Agriculture and Life Sciences, Seoul National University, 2Biomodulation major and Center for Food and Bioconvergence, Seoul National University, 3Laboratory Science Division, International Vaccine Institute, 4 Graduate School of Pharmaceutical Science, Ewha Womans University, 5 Department of Oral Microbiology and Immunology, DRI and BK21 Program, School of Dentistry, Seoul National University, Seoul, Korea Tel: 02-880-4802, E-mail: [email protected] Respiratory syncytial virus (RSV) causes a common respiratory disease, especially for the infants and elderly, whose symptoms are accompanied by a serious bronchiolitis. Up to date, we have no good strategy against RSV infection while it remains a significant cause of death. Indeed poor innate defense mechanisms at initial stage resulted in detrimental consequence in RSV infection, and thus several attempts have been made to improve innate immunity. Biomaterial originated from probiotics would be a good alternative to boost innate immune response owing to their safety and stability. In the present study, mice had been pre-treated with biomaterials and infected with RSV showed less body weight loss and lower viral load than those of control. Mice, with/without RSV infection, administered with biomaterials also displayed changes on the population of innate immune cells, especially significant increase of alveolar macrophages (AMs). Depletion of AMs by using clodronate encapsulated liposomes in these mice, by contrast, resulted in remarkably severe lung morbidity. In addition, bone marrow-derived macrophages (BMMs) and MH-S (alveolar macrophages cell line) stimulated with biomaterials showed advancement of their antiviral ability evidenced by the raise of activation molecules CD40, CD80, CD86 and low number of viral plaques. To elucidate the signaling pathways of biomaterial-induced protective response, we conducted the same setting of experiments using MyD88−/− mice. The results showed the failure of immune cell recruitment with catastrophic disease symptoms including persistence of high viral titer and intensification of histopathology. MyD88−/− BMMs also manifested deficient in antiviral ability by proving high number of plaque compared with wild type BMMs. Taken together, these results demonstrated that biomaterials can increase the action of AMs in the lung and enhance their antiviral activity during the early phase of infection through MyD88-dependent manner. Keywords: Alveolar macrophage, Respiratory syncytial virus, Probiotics-derived biomaterials, Innate immunity, MyD88 signaling The 2014 Fall Conference of the Korean Association of Immunologists 87 Poster Presentation P-117 Innate Immune Response, Infection and Inflammation Imaging of Contact Dynamics of Natural Killer Cells at the Single Cell Level: Identification of Immunocompetent Natural Killer Cells Ji Sung Kim, Yong Guk Kim, Juyoung Kim, Eun Jae Park, Hong Kyung Lee, Hwa Sun Ryu, Minji Pyo, JinBeom Eom, Jin Tae Hong, Youngsoo Kim, Sang-Bae Han* College of Pharmacy, Chungbuk National University, Cheongju, Korea Tel: 043-261-2815, E-mail: [email protected] Natural killer (NK) cells are a type of cytotoxic lymphocyte to eliminate viral infected or transformed cells without antigen-dependent activation. NK cell-mediated killing of target cells is essentially dependent on the direct cell-cell interaction, which has been proved at population levels. However, little is known about contact dynamics of NK cells with target cells at the single cell level. Using time-lapse imaging, we examine how NK cells contact melanoma cells and whether all NK cells are efficient killers of melanoma cells. First, we analyzed contact dynamics of NK cells. Among 1,134 NK cells appeared in 36 movies in time-lapse imaging for 6 h, 121 NK cells (named competent NK, cNK) showed lytic contact with melanoma cells and 158 NK cells (named incompetent NK, iNK) showed non-lytic contact. Although contact numbers of both cell types was similar each other, the duration of contact appeared to be different. cNK cells contacted melanoma cells for 25 min without movement, but iNK cells contacted target cells for 13~14 min with occasional movement. Both cell types moved at average speed of 5~6 um/min before contact and 1.6~2.1 um/min during contact. Next, we analyzed lytic process of melanoma cells, which could be described by two phases: rounding and membrane damage. Immediately after contacting cNK cells, all melanoma cells rounded up and it took 35 min (6-120) having small deviation. Then, melanoma cells lost membrane integrity and it took 103 (6-298) showing big deviation. Together, our data demonstrate that a small fraction of NK cells are cytotoxic to melanoma cells. P-118 Toll like receptor 4 is Critical for Host Resistance Against the Lung Infection of Yersinia pseudotuberculosis in Mice Jin-A Choi1, Yu-Jin Jeong1, Sang-Muk Oh1, Kyung-Bok Lee1, Dong-Jae Kim1, Jong-Hwan Park2* 1 Department of Biochemistry, College of Medicine, Konyang University, Daejeon, 2Department of Laboratory Animal Medicine, College of Veterinary Medicine, Chonnam National University, Gwangju, Korea Tel: 062-530-2834, E-mail: [email protected] Among Yersinia species, Y. enterocolitica, Y. pseudotuberculosis, and Y. pestis are known to be pathogenic to humans. Because of genetic similarities with Y. pestis, Y. pseudotuberculosis has been used to develop systemic infection model of mice via respiratory route, although it causes enteric infection in mammals naturally. However, the model does not entirely mimic the infection of Y. pestis in mice in terms of bacterial spreading. Moreover, host defense mechanism against lung infection of Y. pseudotuberculosis is poorly understood. Therefore, in the present study, we investigated the immunopathological phenomena induced by the lung infection of Y. pseudotuberculosis in WT and TLR4-deficient mice. TLR4 deficiency accelerated lethality by its lung infection in mice. In addition, bacterial loads in the lung, spleen, liver, and blood were much more in TLR4-deficient mice, as compared with WT animals. The levels of cytokines and chemokines in lysates of the lung, spleen, and liver and serum were generally higher in TLR4-deficient mice than WT mice. These finding suggest that TLR4 is essential for the protection of mice against the lung infection of Y. pseudotuberculosis and TLR4 deficient mice may be useful as an animal model of systemic infection for studies of Y. pestis. Keywords: Yerinia pseudotuberculosis, Yersinia pestis, Toll like receptor 4, Lung infection Keywords: NK cells, Dynamics, Imaging, Cytotoxicity P-119 Passively Targeted Intracellular Macromolecule Delivery into Various Immune Cells by Cell Membrane Penetrating Peptide Jung-Ah Lee, Sangho Lim, Won-Ju Kim, Je-Min Choi* Department of Life Science, Hanyang University, Seoul, Korea Tel: 02-2220-4765, E-mail: [email protected] Cell membrane penetrating peptide (CPP) is a short signal peptide that can deliver various cargos including proteins, peptides, DNAs, etc. into the cells. For the last decades, various CPPs have been identified and extensively studied for the application of them on many disease models. However, they have very limited delivery efficiency on primary cells and in vivo, which is a hurdle for the drug development by using CPPs. Here, we used various CPPs including TAT-CPP and novel CPPs such as LPIN, AP, dNP2 to investigate their heterogeneous delivery efficiency depending on various immune cell types and their activation state. Most of CPPs-recombinant proteins were more efficiently delivered into leukemic T cells than primary T cells. In addition, splenic dendritic cells, macrophages showed significantly higher protein uptake efficiency than lymphocytes. Interestingly, the CPP delivery efficiency seems to correlate with cell size and the amount of granules. Furthermore, activated splenic T and B cells showed increased CPP-protein delivery efficiency than naïve cells suggesting CPP has passively targeting property based on cell types and their activation state. We also demonstrated that lipid raft-mediated endocytosis involves intracellular delivery of CPPs-recombinant proteins in activated T cells. Our results provide heterogeneity of CPP mediate protein delivery efficiency on various immune cells and their activation state, which supports valuable information for CPP application strategy. Keywords: Cell membrane penetrating peptide, Primary immune cell, Lipid raft-mediated endocytosis P-120 IL-33-Induced Hematopoietic Stem and Progenitor Cell Mobilization Depends Upon CCR21 Juyang Kim1†, Wonyoung Kim1†, Hongnga T. Le2, 2 2 2 U-jeong Moon , Vuvi G. Tran , Hyun J. Kim , 2 2 3 Quang-Tam Nguyen , Byung-Sam Kim , Jae-Bum Jun , Hong R. Cho1,4†, Byungsuk Kwon1,2†* 1 Biomedical Research Center, Ulsan University Hospital, School of Medicine, University of Ulsan, 2School of Biological Sciences, University of Ulsan, 3Department of Internal Medicine, Ulsan University Hospital, 4 School of Medicine, University of Ulsan, Department of Surgery, Ulsan University Hospital, School of Medicine, University of Ulsan, Ulsan, Korea † These authors contributed equally to this study. Tel: 052-259-1546, E-mail: [email protected] IL-33 has been implicated in the pathogenesis of asthma, atopic allergy, anaphylaxis and other inflammatory diseases by promoting the production of pro-inflammatory cytokines and chemokines or Th2 immune responses. In this study, we analyzed the in vivo effect of IL-33 administration. IL-33 markedly promoted myelopoiesis in the bone marrow (BM) and myeloid cell emigration. Concomitantly, IL-33 induced hematopoietic stem and progenitor cell (HSPC) mobilization and extramedullar hematopoiesis. HSPC mobilization was mediated mainly through increased levels of CCL7 produced by vascular endothelial cells in response to IL-33. In vivo treatment of IL-33 rapidly induced phosphorylation of ERK, JNK and p38 and inhibition of these signaling molecules completely blocked the production of CCL7 induced by IL-33. Consistently, inhibitor of CCR2 markedly reduced IL-33-mediated HSPC mobilization in vivo and migration of HSPC in response to CCL7 in vitro. IL-33-mobilized HSPCs were capable of homing to and of long-term reconstitution in the BM of irradiated recipients. Immune cells derived from these recipients had normal anti-fungal activity. The ability of IL-33 to promote migration of HSPCs and myeloid cells into the periphery and to regulate their anti-fungal activity represents a previously unrecognized role of IL-33 in innate immunity. These properties of IL-33 have clinical implications in hematopoietic stem cell transplantation. Keywords: IL-33, CCR2, Hematopoietic stem and progenitor cell 88 The 2014 Fall Conference of the Korean Association of Immunologists Innate Immune Response, Infection and Inflammation P-121 CXCL10-CXCR3 Network Regulates Natural Killer Cell Migration to B16F10 Melanoma Cells Juyoung Kim, Ji Sung Kim, Yong Guk Kim, Eun Jae Park, Hong Kyung Lee, Hwa Sun Ryu, Minji Pyo, JinBeom Eom, Jin Tae Hong, Youngsoo Kim, Sang-Bae Han* College of Pharmacy, Chungbuk National University, Cheongju, Korea Tel: 043-261-2815, E-mail: [email protected] Natural killer (NK) cells play critical roles in host immunity against cancer. NK cells directly kill target cells through several mechanisms. Main event of NK cell cytotoxicity is direct cell-cell contact with target cells. Although NK cell migration to the cancer cell is essential for direct cell-cell interaction, how tumor cells regulate NK cell migration is not fully defined. Using time-lapse imaging, we studied whether tumor cell-derived chemokines affect migration and cytotoxicity of NK cells. First, we increased chemokine production by cancer cells and examined NK cell migration. We treated B16F10 cells with IFN-γ, which resulted in high production of CXCL10 compared with parent B16F10 cells. IL-2-acitvated NK cells expressed chemokine receptors such as CCR2, CCR5 and CXCR3. When cultured with IFN-γ-treated B16F10 cells, NK cells showed faster speed and better migration of NK cells. Subsequently, NK cells showed better killing to IFN-g-treated B16F10 cells than parent cells. These data suggest that NK cells migrate well toward cancer cells highly producing CXCL10. Second, we used CXCR3−/− NK cells, which could not respond to CXCL10. CXCR3+/+ NK cells migrated well toward IFN-g-treated B16F10 cells and moved at a speed of 7.4 um/min. However, CXCR3−/− NK cells migrated poorly toward IFN-γ-treated B16F10 cells and moved at 6.4 um/min. Subsequently, CXCR3−/− NK cells killed target cells much less than CXCR3+/+ NK cells did. Together, our data suggest that tumor-derived chemokine CXCL10 upregulates moving speed and chemotactic migration of NK cells, which increases the chance to contact and kill target cells. P-122 Poster Presentation Yersinia pestis LPS Induces Nitric Oxide Production in Mouse Macrophages through Toll-Like Receptor 4 Ok-kyu Park, Sang Yoon Choi, Gi-eun Rhie, Jun Ho Jeon* Division of High-risk Pathogen Reserch, Center for Infection Diseases, National Institute of Health, Cheongju, Korea Tel: 043-719-8287, E-mail: [email protected] Yersinia pestis (Y. pestis), the etiological agent of plague, is a highly virulent Gram-negative, rod-shaped, facultative anaerobic bacterium that causes a life-threatening infectious disease to humans and other animals. In the present study, we purified lipopolysaccharide (LPS) from Y. pestis and investigated the effect of nitric oxide (NO) production in a mouse macrophage cell-line, RAW264.7 cells. Upon treatment of cells with Y. pestis LPS, NO production was increased in a dose-dependent manner. Furthermore, this NO production appeared to be due to increase in the inducible nitric oxide synthase (iNOS) mRNA and protein levels. Y. pestis LPS increased NO production in mouse bone-marrow-derived macrophages (BMDMs) and this enhancement was completely abrogated in BMDMs from Toll-like receptor (TLR) 4 mutant or MyD88-deficient mice. Western blot analysis showed that Y. pestis LPS induced phosphorylation of MAP kinases including ERK, JNK, and p38. Furthermore Y. pestis LPS-induced NO production was decreased by selective inhibitors of all three MAP kinases. Y. pestis LPS induced degradation of IκB-α which is an inhibitory molecule of NF-κB as well as phosphorylation of Akt. Concomitantly, Y. pestis LPS-induced NO production was significantly attenuated by inhibitors of PI3 kinases and NF-κB. Collectively, these results suggest that Y. pestis LPS-induced NO production requires activation of TLR4, MyD88, and subsequent activation of MAP kinases and PI3 kinases leading to the activation of the transcription factor NF-κB in mouse macrophages. Keywords: Yersinia pestis, lipopolysaccharide, nitric oxide, TLR4 Keywords: NK cells, CXCR3, CXCL10, Migration P-123 A Recombinant Fusion Protein F1-V Provides Protection Against a Lethal Yersinia pestis Challenge in Mouse Model Sang-Yoon Choi, Ok Kyu Park, Gi-eun Rhie, Jun-Ho Jeon* Division of high-risk pathogen research, Center for Infectious Diseases and Prevention, Korea National Institute of Health, Osong Health Technology Administration Complex, Cheonju, Korea Tel: 043-719-8298, E-mail: [email protected] The causative organism of plague is known to Yersinia pestis. Y.pestis is a Gram-negative, rod-shaped that causes bubonic and pneumonic plague and has a potential for deliberate use as a bioterrorism or warfare. Currently, there is no FDA approved plague vaccine available. In this study, the recombinant protein F1-V was evaluated not only the potential vaccine candidate but also immunogenicity and protective efficacy. The IgG antibody titers of immunized group (rF1, rV, rF1-V and rV-F1) were significantly higher than PBS and Alum group. Also, antibody titers of fusion proteins rF1-V were higher than rF1, rV and V-F1 protein in immunized mice. And, anti-rF1-V and anti-rV-F1 was 3-4 folds higher serum igG titer than anti-F1, respectively. The survival percentage seen among the rF1-V immunized mice group (60%) was significant (P <0.05) when compared with control (PBS and Alum) or other groups immunized with rF1, rV and rV-F1. Taken together, the fusion protein rF1-V could be used as a potential candidate for the subunit vaccine for prevention of Y. pestis. Keywords: Y. pestis, Plague, Fusion protein, F1-V P-124 mTOR-Independent Induction of Autophagy Preferentially Modulates Monosodium Urate Crystal-Induced Interleukin-1β Production in Humans Yeon-Ho Chung, Won-Woo Lee* Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul, Korea Tel: 02-740-8303, E-mail: [email protected] Monosodium urate (MSU) crystal-induced interleukin-1β (IL-1β) release from innate immune cells is a critical factor in the pathogenesis of gouty inflammation. Activation of the NLRP3 inflammasome by MSU crystal is required for the release of mature IL-1β. Autophagy is a conserved cellular processes for maintaining homeostasis by degrading damaged organelle and long-lived proteins and has the potential to impact the progress of inflammatory diseases. However, the role of autophagy in regulating gouty inflammation remains unclear. In this study, we investigated the putative regulatory roles of autophagy affecting MSU crystal-induced IL-1β production in human monocytic THP-1 cells. Immunoblotting analysis revealed that an autophagy inducer, rapamycin and trehalose, increased endogenous microtubule-associated protein light chain 3-II (LC3-II) which is used as a marker for autophagosome. Confocal microscopic analyses also showed that autophagy inducers enhanced the number and the intensity of LC3-II puncta. However, inhibition of autophagy with 3-methyladenine (3-MA) promoted secretion of IL-1β by MSU crystal treatment. Although rapamycin (mTOR-dependent autophagy inducer) does not affect the release of mature IL-1β, induction of autophagy with trehalose (mTOR-independent autophagy inducer) decreased intracellular pro-IL-1βlevel and reduced release of mature IL-1βfrom MSU crystal treated THP-1 cells. These results suggest that autophagy can modulate inflammatory responses in MSU crystal-induced gouty inflammation. Keywords: MSU crystal, Gout, IL-1β, Autophagy The 2014 Fall Conference of the Korean Association of Immunologists 89 Poster Presentation P-125 Innate Immune Response, Infection and Inflammation Characterization of Human Natural Killer Cell Subsets According to the Expression of CD49a and CD49b Integrins Youkyong Song, Joo Chun Yoon* Department of Microbiology, Ewha Womans University School of Medicine, Seoul, Korea Tel: 02-2650-5740, E-mail: [email protected] Background: Natural killer (NK) cells are lymphocytes of the innate immunity and participate in antiviral and antitumor immune responses. Integrins CD49a and CD49b (DX5) on NK cells mediate adhesion to other cells or extracellular − + matrix. NK cells in the mouse blood are CD49a DX5 , whereas liver-resident + − NK cells have another subset which is CD49a DX5 . We therefore investigated whether primary human NK cells are also separated into CD49a/bbased subsets and identified functional characteristics of each subset. Methods: Peripheral blood mononuclear cells were isolated from whole blood of healthy human donors. CD49a/b expression, degranulation, IFN-γ production, and TRAIL expression were analyzed by flow cytometry. Results: Primary human NK cells expressed CD49a and CD49b and could be + − − + + + divided into four subsets: CD49a CD49b , CD49a CD49b , CD49a CD49b , − − and CD49a CD49b . The frequency of human NK cell subsets, however, differed from that of mouse NK cell subsets. Interestingly, the proportion of + − CD49a CD49b NK cells increased after overnight ex vivo culture. In particbright dim NK cells expressed more CD49a than CD56 NK cells. In ular, CD56 − + contrast, the frequency of CD49a CD49b NK cells decreased after ex vivo culture. Degranulation capacity of NK cells increased after the culture and it + − was largely due to increase of CD49a CD49b NK cells. IFN-γ production was not affected by CD49a/b expression. TRAIL expression was positive on + + − + CD49a CD49b NK cells and CD49a CD49b NK cells in the resting state. Upon stimulation with IFN-α-2b, however, TRAIL expression was sig+ + nificantly elevated on the surface of all NK cell subsets except CD49a CD49b + NK cells, of which more than 70% are TRAIL even before IFN-α-2b treatment. Conclusion: Human NK cells were divided into four subsets based on CD49a/b expression. While degranulation capacity was related to CD49a expression, it appeared that TRAIL expression in resting NK cells was associated with CD49b expression. Keywords: Human natural killer cell, CD49a, CD49b, Subset, Function P-127 Deficiency of Melanoma Differentiation-associated Protein 5 (MDA5) Results in Exacerbated Chronic Postviral Lung Inflammation 1 1 1 Won-keun Kim , Deepika Jain , Melissa D. Sanchez , 2 3 Cynthia J. Koziol-White , Krystal Matthews , Moyar Q. Ge2, Angela Haczku2, Reynold A. Panettieri, Jr.2, Matthew 3 1 B. Frieman , Carolina B. Lopez * 1 Department of Pathobiology, School of Veterinary Medicine, University 2 of Pennsylvania, Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, 3Department of Microbiology and Immunology, School of Medicine, University of Maryland at Baltimore, Maryland, USA Tel: 02-920-6354, E-mail: [email protected] Respiratory viral infection can lead to the establishment of chronic lung inflammatory diseases including chronic obstructive pulmonary disease (COPD). Understanding the early innate immune mechanisms that participate in the development of chronic postviral lung disease may be critical for therapeutic intervention. However, little is known of the impact of the innate immune system on the development of long-term lung disease. The melanoma differentiation-associated gene 5 (MDA5) is a germ line encoded pattern recognition receptor (PRR) that plays a role in detecting molecular motifs present in viral oligonucleotides. Sendai virus (SeV) causes bronchiolitis and persistent airway hyperreactivity in mice, providing an accepted experimental model for virus-induced chronic lung disease. Here we show MDA5 deficiency results in a distinct profile of chemokines and cytokines that associated with acute lung neutropenia, enhanced accumulation of alternatively activated macrophages, and sustained lung inflammation upon infection with SeV while viral loads remain unchanged. Remarkably, the biased acute inflammatory response of MDA5 deficiency elicits an enhanced chronic lung inflammation, epithelial cell hyperplasia, airway hyperreactivity, and diminished blood oxygen saturation. Overall, these studies demonstrate that the intracellular viral sensor molecule MDA5 modulates the development of chronic lung inflammation by regulating the early inflammatory response in the lung. Keywords: Respiratory viral infection, Innate immune response, MDA5, Alternatively activated macrophage, Chronic Lung Inflammatory response 90 P-126 Distinct Proinflammatory Properties of Peripheral Monocytes in Patients with End-Stage Renal Disease (ESRD) Yuri Hwang1, Jiyeon Jang1, Sungha Park2, Won-Woo Lee1,3* 1 Department of Microbiology and Immunology, Seoul National University College of Medicine, 2Division of Cardiology, Severance Cardiovascular Hospital and Cardiovascular Research Institute, Yonsei University College of Medicine, 3Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, Korea Tel: 02-740-8303, E-mail: [email protected] Loss of renal function is closely related to proinflammatoy milieu and a concomitant immune dysfunction in humans. In patients with end-stage renal disease (ESRD), uremia-associated chronic inflammation has been known to be involved with an increased risk of cardiovascular disease (CVD) as well as aberrant immune responses, especially innate immune responses. Monocytes are known for the central driver of vascular inflammation in CVD. Human monocytes are heterogeneous and can be classified into three distinct subsets based on CD14 and CD16 expression. Although accumulating evidence suggests distinct functions of monocyte subsets in inflammatory conditions, their pathogenic roles in ESRD remain unclear. Here we investigated functional characteristics of monocyte derived from peripheral blood in ESRD patients. In our study, the frequency of proinflammatory CD14+CD16++ monocytes was significantly increased in ESRD patients at the expense of diminished frequency of CD14++CD16- monocytes when compared with that in age-matched HCs. It has been reported that these CD14+CD16++ monocytes produced a large amount of various proinflammatory cytokines upon stimulation and those cytokines might be involved in various pathogenic inflammatory disorders. To identify gene expression signature of monocytes in patients with ESRD, we compared a global transcriptome of monocytes between ESRD patients and age-matched HCs using microarray analysis. Our data showed that 3,976 genes were differentially expressed in monocytes of ESRD patients compared with those of age-matched HCs. In particular, inflammatory chemokines, CCL3 and CXCL8 (IL-8) and pro-inflammatory cytokine, IL-1β gene expression were markedly elevated in ESRD monocytes. Given that these inflammatory chemokines play a critical role for migration of inflammatory leukocytes into the damaged in CVD, these results suggest that proinflammatory milieu by monocytes might be associated with an increased risk of CVD in ESRD patients. Keywords: Monocyte, End-stage renal disease (ESRD), Uremia, Cardiovascular disease (CVD), Microarray analysis, Inflammation P-128 Purification and Functional Characterization of EsxGH Complex Antigen of Mycobacterium abscessus Isaac Kim1,2, Loi T. Nguyen3, Joy G. Lee1,2, Myung Hee 3 1,2 Kim , Eun-Kyeong Jo * 1 Department of Microbiology, 2Infection Signaling Network Research 3 Center, Chungnam National University School of Medicine, Infection and Immunity Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea Tel: 042-580-8243, E-mail: [email protected] Mycobacterium abscessus (Mabc) is a nontuberculous and a rapidly growing mycobacterium which often causes invasive lung diseases and opportunistic infection in immunocompromised patients. Although the whole genome sequences of different Mabc strains were identified several years ago, the characterization of each gene/protein is just beginning. Mycobacterial ESX clusters have been known to be involved in the virulence of pathogenic mycobacteria, especially M. tuberculosis. However, little is known about the identification and functional characterization of Mabc Esx proteins in innate immune cells. In this study, we purified the EsxGH complex proteins to investigate its functional and immunological properties. The N-terminal hexahistidine-tagged EsxG protein and the N-terminal GST-fused EsxH protein were expressed in E. coli BL21 (DE3) and E. coli C43 (DE3) systems, respectively. The expressed EsxG and EsxH proteins were then purified by Ni-NTA and GST affinity chromatographiesy, respectively. EsxH was further purified by treatment with rTEV protease to remove GST and additional GST affinity chromatography. The complex of the purified EsxG and EsxH proteins was formed spontaneously upon mixing and subjected to size-exclusion chromatography for further purification. The purified EsxGH complex proteins triggered the macrophage inflammatory activities in murine bone marrow-derived macrophages (BMDMs). We found that Mabc EsxGH complex proteins robustly induced inflammatory cytokine gene expression in BMDMs. Together, these data suggest that the Mabc EsxGH complex plays an important role in the activation of inflammatory responses in innate immune cells. Keywords: M. abscessus, Esx The 2014 Fall Conference of the Korean Association of Immunologists Innate Immune Response, Infection and Inflammation P-129 MicroRNA-125a Inhibits Autophagy Activation and Antimicrobial Responses during Mycobacterial Infection P-130 Poster Presentation Insufficiency of Vitamin C Induces a Defect on the Fetal Growth and Maintenance of Pregnancy in Gulo(−/−) Mice Jin Kyung Kim1,2, Jae-Min Yuk2,3, Suyeon Kim1,3, Tae Sung Kim1,3, Hye-Mi Lee1,3, Hyo Sun Jin1,3, Eun-Kyeong Jo1,3,4* Hyemin Kim, Yejin Kim, Jiwon Choi, Mirim Jang, Jiyea Choi, Junman Hong, Hyunwoo Kim, Young-il Hwang, Jae Seung Kang, Wang Jae Lee* 1 Department of Anatomy, Seoul National University College of Medicine, Seoul, Korea Tel: 02-740-8202, E-mail: [email protected] 2 3 Department of Microbiology, Department of Infection Biology, Infection Signaling Network Research Center, 4Research Institute for Medical Sciences, Chungnam National University School of Medicine, Daejeon, Korea Tel: 042-580-8358, E-mail: [email protected] MicroRNAs (miRNAs) are small noncoding RNA molecules (~22 nucleotides in length) that regulate gene expression at the post-transcriptional level. Through binding to the 3′-untranslated regions (UTRs) of target messenger RNAs (mRNAs), miRNAs typically lead to suppression of protein translation or mRNA degradation. Autophagy is a lysosome-mediated catabolic process that is important for the degradation of unwanted cytoplasmic cargos. Increasing data reveal that autophagy plays a key role in activating the antimicrobial host defense against Mycobacterium tuberculosis (Mtb). Although the pathways associated with autophagy must be tightly regulated at a post-transcriptional level, the contribution of miRNAs to this process, including whether they specifically influence the activation of macrophage autophagy during Mtb infection, is largely unknown. In this study, we investigated whether the induction of miRNA-125a-3p (miR125a) in response to Mtb infection is involved in targeting autophagy-related genes. We thus infected RAW264.7 cells and murine bone marrow-derived macrophages (BMDMs) with Mtb, and measured the expression level of miR125a using quantitative RT-PCR analysis. We found that Mtb-infected BMDMs showed a gradual increase in expression of miR125a in a moi-dependent manner. In addition, Mtb infection increased miR125a expression in both BMDMs and RAW264.7 cells. We further identified specific mRNA targets with miR125a-binding sites by a bioinformatic analysis. Interestingly, UVRAG mRNA was predicted to be a potential mRNA target of miR125a, through interaction with a complementary sequence in the 3′ UTR. Using 3'UTR luciferase reporter assays, we further demonstrated the miR125a-dependent suppression of luciferase activity in RAW264.7 cells expressing the wild-type UVRAG 3'UTR reporter. Together, these results indicate that miR125a post-transcriptionally inhibits UVRAG expression through a direct interaction with its 3'UTR binding site. Developing fetus is particularly susceptible to vitamin C deficiency because rapid growth and immature antioxidant system. So, we investigated the effect of vitamin C on the fetal development using Gulo(-/-) mice. When maternal Gulo(-/-) mice was depleted of vitamin C for 2 weeks during pregnancy, the serum level was vitamin C was half of vitamin C-sufficient Gulo(-/-) mice or wild-type (WT) mice. The number and body weight of fetus was reduced, and the concentration of vitamin C in the amniotic fluid was significantly decreased in the vitamin C-insufficient Gulo(-/-) mice. Moreover, Gulo(-/-) mice showed a loose integrity, an increased expression of matrix metalloproteinase 9 (MMP-9), and a decreased vascular permeability in the placenta. Also, the production of progesterone, a hormone for maintaining pregnancy, was considerably reduced. Therefore, vitamin C insufficiency during gestation could disturb the fetal growth and maintenance of pregnancy. Keywords: Vitamin C insufficiency, Gulo(-/-) mice, Fetal growth, Pregnancy Keywords: MicroRNA, Autophagy, Mycobacteria, UVRAG, Macrophages P-131 The Effect of Yersinia pestis LPS on Nitric Oxide (NO) Production in RAW264.7 Cells Ok-kyu Park, Sang Yoon Choi, Gi-eun Rhie, Jun Ho Jeon* Division of High-risk Pathogen Reserch, Center for Infection Diseases, National Institute of Health, Cheongju, Korea Tel: 043-719-8287, E-mail: [email protected] Yersinia pestis (Y. pestis), the etiological agent of plague, is a highly virulent Gram-negative, rod-shaped, facultative anaerobic bacterium that causes a life-threatening infectious disease to humans and other animals. Also, role of the lipopolysaccharide (LPS) is one of the major pathogenicity factors in Y. pestis. Our data showed that on Nitric oxide(NO) production in Y. pestis lipopolysaccharides(LPS)-stimulated RAW264.7 cells. Treatment of RAW264.7 cells with 27oC or 37oC Y. pestis LPS induced NO production. This induction of NO was completely abrogated in mouse bone marrow-derived macrophage cells(BMDMs) from TLR4 mutant mice, C3H/HeJ. Y. pestis LPS induced phosphorylation of all three MAP kinases including ERK, JNK, and p38. LPS-induced NO production was decreased by inhibitors of all three MAP kinases. In addition, Y. pestis LPS induced a NF-κB and PI3 kinases signaling pathways. LPS-induced NO production was decreased by inhibitors of a NF-κB and PI3 kinases. These results suggest that TLR4 and MYD88- dependent signaling pathways are involved in Y. pestis LPS-induced NO production. Furthermore Y. pestis LPS-induced NO production occurs inflammatory responses partly via TLR4-mediated MAP kinases, a transcription factor NF-κB and PI3 kinases pathways in RAW264.7 cells. P-132 Salmonella Typhimurium Infection Increases G-CSF Expression in Macrophages Jun Beom Park, Tae Sung Kim* Department of Life Sciences, Korea University, Seoul, Korea Tel: 02-3290-3920, E-mail: [email protected] In infection with Salmonella Typhimurium, neutrophil has a crucial role for ingesting microorganisms or their particles. Granulocyte colony stimulating factor (G-CSF) regulates the generation of neutrophils in the bone marrow and their release into the blood. In infection with Listeria monocytogenes, it has been known that G-CSF from infection affects the generation of neutrophils in the bone marrow via serum. In infection with Salmonella Typhimurium, however, it is not well known about the induction of G-CSF in macrophages and the mechanism of neutrophil generation by G-CSF. In this study, we investigated whether RAW264.7, a murine macrophage cell line, induced the expression of G-CSF by the infection with Salmonella Typhimurium. Infection with Salmonella Typhimurium increased significantly G-CSF expression in RAW264.7 in time- and MOI-dependent manners. Consequently G-CSF from macrophages infected with Salmonella Typhimurium may play a role in bacterial clearance through generation of neutrophils. Regulatory molecules in Salmonella Typhimurium which are involved in G-CSF expression are under investigation by using its gene mutants in macrophage culture systems. Keywords: Salmonella Typhimurium, G-csf, Murine macrophage, Neutrophil Keywords: Yersinia pestis, LPS, Nitric oxide production, TLR4 The 2014 Fall Conference of the Korean Association of Immunologists 91 Poster Presentation P-133 Innate Immune Response, Infection and Inflammation Differentially Oxidized Low-Density Lipoprotein Affects Macrophage Phenotype in Human Monocytic Cells Jin-Won Seo1,2, Eun-Jeong Yang1,3, In-Hong Choi1,2,3* 1 Department of Microbiology, 2Brain Korea 21 PLUS Project for Medical 3 Science, Yonsei University College of Medicine and Institute for Immunology and Immunological Disease, Seoul, Korea Tel: 02-2228-1821, E-mail: [email protected] Atherosclerosis is a chronic inflammatory disease of the arterial wall. Many studies have shown that retention of low-density lipoprotein (LDL) and recruitment of monocytes into subendothelial space are critical early events in atherosclerotic development. Accumulated LDL in the arterial wall is undergone oxidative modification according to a variety of oxidizing agents and the exposure duration. For these reasons, different degrees of oxidized LDL exist in the atherosclerotic plaque. In this study, we examined the effect of differentially oxidized LDL in differentiation of human monocytic cells. Human monocytic THP-1 cells and human blood monocytes were incubated with two different oxidized LDL (low-oxLDL and high-oxLDL). First, we observed the effect of different oxidized LDL for morphological changes. Human blood monocytes treated with low-oxLDL showed spindle shape, however, the cells treated with high-oxLDL showed round shape. The expression of classically-activated macrophage marker (CD86) was increased in THP-1 cells treated with low-oxLDL, but, the expression of alternatively-activated macrophage marker (CD206) was increased in THP-1 cells treated with high-oxLDL. Also the formation of foam cells was significantly increased in THP-1 cells treated with high-oxLDL compared to the cells treated with low-oxLDL. Intriguingly, we found that granularity and autofluorescence of THP-1 cells treated with oxLDL were increased in oxidation dependent manner. Although pro- and anti-inflammatory cytokines were not detected the production of interleukin-8 was significantly increased in low-oxLDL treated THP-1 cells compared to cells treated with more highly oxidized LDL. These results suggest that the degree of oxidation of LDL influences differentiation of human monocytic cells. THP-1 cells treated with low-oxLDL may differentiate into classically-activate macrophages, while THP-1 cells treated with high-oxLDL may differentiate into alternatively-activate macrophages. P-134 Transgelin2 Regulates Macrophage Migration In The Inflammatory Condition Hyun-Su Lee, Bo-Ra Na, Hye-Ran Kim, Min-Sung Kwon, Chang-Duk Jun* School of Life Sciences, Immune Synapse Research Center and Cell Dynamics Research Center, GIST, Gwangju, Korea Tel: 062-715-2567, E-mail: [email protected] Transgelin2 (TAGLN2) is an actin binding molecule while no specific actions are reported. Many actin regulating proteins have been elucidated their functions in macrophages in relation to the actin dynamics such as phagocytosis, adhesion and migration. However, function of TAGLN2 in macrophages is largely unknown. In this study, we found that TAGLN2 is modestly expressed in macrophages but dramatically up-regulated in response to the LPS stimulation. Pharmacologic investigation demonstrated that TAGLN2 expression is controlled by the NF-κB, a major transcription factor responding to the LPS. In consistence with this, analysis of promoter region of TAGLN2 gene contained the NF-κB region, suggesting that TAGLN2 is a NF-κB responsible gene. TAGLN2-deficient macrophages (TAGLN2−/−) showed reduced F-actin contents in activated condition and attenuated FcR-mediated phagocytic activity. −/− Besides, TAGLN2 macrophages indicated impaired migration activity in LPS/fMLP-stimulated condition. We are now going to identify the molecular mechanisms implicated in TAGLN2 action in inflammatory condition. Keywords: Transgelin2, Macrophage, Migration, Inflammation, Actin polymerization Keywords: Atherosclerosis, Oxidized Low-density lipoprotein (OxLDL), Monocyte, Macrophage P-135 Alternatively Activated Macrophage Facilitates Recovery From Intracerebral Hemorrhage Yong Ho Jang, Hyunjung Min, Sung Joong Lee* Department of Neurobiology and Physiology, Dental Research Institute, School of Dentistry, Seoul National University, Seoul, Korea Tel: 02-740-8649, E-mail: [email protected] Intracerebral hemorrhage (ICH) is one of the major causes of stroke, constituting 15% to 20% of all stroke cases. Recently, it has been reported that the function of macrophages is distinct depending on their activation type; M1, or classical activation, leads to pro-inflammatory responses, while M2, or alternative activation, contributes to tissue repair or healing. Since the function of brain-infiltrating macrophages after ICH has not been elucidated, we characterized the phenotype of macrophages infiltrating brain parenchyma and investigated their function in ICH. We found that CX3CR1+ myeloid cell number increased at perihematoma sites following ICH injury and the mRNA expression of Arginase-1 (an M2 marker) was upregulated in the ICH-injured brain, while iNOS (an M1 marker) expression was not significantly altered. In our study using flow cytometry we found that mannose receptor (an M2 marker)-expressing macrophages are further augmented at a more delayed time point after ICH. In the macrophage-depleted mice the ICH-induced brain lesions were more sustained compared to those of control mice, indicating a putative role of macrophages in tissue healing and recovery. The M2 polarization of the brain-infiltrating macrophages suggested that the brain microenvironment around macrophages may affect macrophage activation. Upon co-culture with MBMG, the number of mannose receptor-positive BMDMs was significantly increased, which suggests that glia cells, the most abundant cell type in the CNS, can induce macrophage M2 polarization. Furthermore, treatment with MBMG conditioned media increased mannose receptor-expressing BMDMs, as well as implicated MBMG-derived soluble factors in the macrophage M2 polarization. Taken together, our data suggest that brain-infiltrating macrophages, after ICH, are differentiated to the M2 phenotype by brain glial cells, and thereby contribute to the recovery from ICH injury. Keywords: Intracerebral hemorrhage, Neuroinflammation, Macrophage polarization, M2 macrophage, Glia 92 P-136 Association of Toll-Like Receptor 10 Polymorphisms with Autoimmune Thyroid Disease in Korean Children Jung-Pil Jang1, In-Cheol Baek1, Eun-Jeong Choi2, 3 1,2 Won-Kyoung Cho , Tai-Gyu Kim * 1 Department of Microbiology and immunology, College of Medicine, The 2 Catholic University of Korea, Catholic Hematopoietic Stem Cell Bank, College of Medicine, The Catholic University of Korea, 3Department of Pediatrics, College of Medicine, The Catholic University of Korea Tel: 02-2258-7341, E-mail: [email protected] The Toll-like receptors (TLRs) are germline-encoded receptors that play an essential role in initiating the immune response against pathogens. In this study, we assess the association of TLR polymorphism with autoimmune thyroid disease (AITD) in Korean children. We defined the polymorphism of TLR10 gene (rs4129009, rs11096956 and rs10004195) in 85 Korean AITD (Graves disease [GD]=50, Hashimoto disease [HD]=35; thyroid-associated ophthalmopathy [TAO]=23, non-TAO=62; M=16 , F=69; mean age=12.9±3.1 years ) and 183 healthy control subjects. In patients with AITD, the frequencies of the TLR10 rs10004195 TT genotype (OR=2.0, cP=0.05) and T allele (OR=2.7, cP=0.01) were higher than healthy controls, whereas the TLR10 rs4129009 GG genotype (OR=0.2, cP=0.01) and rs10004195 AA genotype (OR=0.4, cP=0.02) and A allele (OR=0.5, cP=0.03) showed lower frequencies. The TLR10 rs11096956 did not show any significant association. These significant associations were also found in non-thyroid associated ophthalmopathy (TAO) group whereas not in TAO. Patients with GD showed the lower frequency of the TLR10 rs10004195 AA genotype (OR=0.4, cP=0.02). The haplotype (AGT) frequency of TLR10 rs4129009, rs11096956 and rs10004195 was higher in AITD than healthy controls (OR=0.3, cP=0.01). Our results suggest that TLR10 polymorphisms may contribute to the pathogenesis of AITD. Keywords: Autoimmune thyroid disease, Toll-Like Receptor 10, Graves disease, Thyroid associated ophthalmopathy, Innate immunity The 2014 Fall Conference of the Korean Association of Immunologists Innate Immune Response, Infection and Inflammation P-137 The Bactericidal Effect of Lysophosphatidylcholin in Salmonella Typhimurim-Infected Macrophages Wan-Gi Hong1, Yu-Jin Jung1,2* 1 BIT Medical Convergence Graduate Program, 2Department of Biological Sciences, Kangwon National University, Chuncheon, Korea Tel: 033-250-8533, E-mail: [email protected] Salmonella enteric serovar Typhimurium (S. Typhimurium) is a intracellular pathogen that infects macrophages and survives in phagosome by avoiding phago-lysosome fusion system. Phago-lysosome fusion system is associated with Rab GTPases proteins and phagosomes sequentially acquire different Rab GTPase proteins during maturation and eventually fuse with acidic lysosomes. Lysophosphatidylcholine (LPC) is known as a major component of oxidized low density lipoprotein but its immunological or pharmacological function was not defined well. Recent studies have shown that LPC enhances the bactericidal activity in neutrophils in animal sepsis model. However, the effect of LPC remains incompletely understood and controversial, as there are several reports that LPC has either stimulatory or inhibitory roles on immune response. In this study, we examined whether LPC would reveal bactericidal effects against intracellular bacteria, S. typhimurim in mouse macrophages. When RAW264.7 cells were treated with LPC after infection of S. typhimurium, the bacterial burden was significantly decreased, as compared with S. typhiumurium-infected cells. To identify the cellular mechanism of increased bactericidal effect of LPC, the expression level of the molecules related with phagosomal maturation were detected. When the expression of Rab5 and Rab7 were measured at the transcriptional and translational levels, both were elevated in LPC-stimulated cells during S. typhimurium infection. The ratio of co-localization of intracellular S. typhimurium with various phagosomal marutaion markers, such as Rab5, Rab7, EEA1 and LAMP-1, increased in LPC-treated cells. In addition, treatment of LPC triggered the increased phosphorylation of IκBα, whereas the expression of phosphorylated MEK, ERK and p38 unchanged during the infection. These findings revealed that LPC would be developed as a therapeutic agent to modulate S. Typhimurium-killing activity in macrophages by enhancing phagosomal maturation. Keywords: LPC (Lysophosphatidylcholine), Phagosomal maturation, Macrophage, Salmonella typhimurium P-139 LPS-Modified Outer Membrane Vesicle (fmOMV) Adjuvant Provides Protective Innate Immunity Against Influenza Infection Prior to the Induction of Vaccine-Induced Adaptive Immune Response 1 1 1 Eun-Hye Bae , Tae-Young Lee , Sang-Ho Lee , Min-Joo Yeom1,2, Woon-Seong Na1,2, Chang-Ung Kim1,3, Dae-Sub 1,2 1,2 1 Song , Doo-Jin Kim *, Sang-Hyun Kim * 1 Viral Infectious Disease Research Center, Korea Research Institute of 2 Bioscience and Biotechnology (KRIBB), Division of Biosystems & Bioengineering, University of Science and Technology (UST), 3Department of Biochemistry, Chungnam National University, Daejeon, Korea Tel: 042-879-8276, E-mail: [email protected] Outer membrane (OM) vesicles, naturally produced by Gram-negative bacteria, have important roles in bacterial pathogenesis. It is also well known that OM vesicles can stimulate the host innate immune system through the activation of toll like receptor (TLR) signaling pathways. Previous studies have revealed that TLR agonists can trigger the innate immune response in the lung and protect against influenza infection. In this study, we investigated the immunostimulatory effect of LPS-modified OM vesicle (fmOMV) in mice, as well as the efficacy of fmOMV as an antiviral agent against influenza virus infection. Intranasal administration of fmOMV dramatically enhanced the survival rate against lethal influenza A virus challenge. The protective immunity maintained for a week. More importantly, pre-treatment of fmOMV conferred broad protection against different influenza virus strains, while the vaccination provided only strain-specific immunity. fmOMV also significantly increased the level of proinflammatory cytokines such as IL-6, RANTES, IL-1β, IFN-γ and TNF-α in the airway lumen, and the neutrophil and macrophages recruitment into the lungs. These data suggest that fmOMV induces robust activation of innate immunity in the lungs and can be used as an effective antiviral agent against influenza viruses. Keywords: OMV, Innate immunity, Toll like receptor, Antiviral agent P-138 Poster Presentation Mycobacterium tuberculosis Induces Immune Response of Murine Pleural Mesothelial Cells via Toll like Receptor 2-Dependent Pathway Eun-Ha Hwang1, Tae-Hyoun Kim2, Sang-Muk Oh1, Kyung-Bok Lee1, Dong-Jae Kim1, Sung-Jae Shin3, Jong-Hwan Park4* 1 Department of Biochemistry, College of medicine, Konyang University, Daejeon, 2Department of Laboratory Animal Medicine, College of Veterinary Medicine, Seoul National University, Seoul, 3Department of Microbiology, Institute for Immunology and Immunological Diseases, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, 4Department of Laboratory Animal Medicine, College of Veterinary Medicine, Chonnam National University, Gwangju, Korea Tel: 062-530-2834, E-mail: [email protected] Pleural effusion is characterized by the accumulation of fluid and inflammatory cells in the pleural space, which occurs through a mechanism that is still not completely understood. Mycobacterium tuberculosis is a causative agent leading pleural effusion. Pleural mesothelial cells (PMCs) are specialized epithelial cells that line the internal organs and body wall in pleural cavity so that play an important role in pleural inflammation. Toll like receptors are expressed in various cell type including mesothelial cells and play a key role in Mycobacterium infection. In the present study, we investigated direct immune responses of PMCs against BCG and M. tuberculosis and the role of TLR2 in such responses. BCG and M. tuberculosis increased the production of IL-6, CXCL1, and CCL2 in WT PMCs, which was impaired in TLR2-deficient cells. In addition, the activation of NF-κB and MAPKs induced by BCG and M. tuberculosis was delayed in TLR2-deficient PMCs, as compared with WT cells. TLR2 deficiency led to the decrease of Nitric oxide (NO) production and delay of the gene expression of iNOS in PMCs. TLR2 was also essential for optimal expression of ICAM-1 and VCAM-1, cellular adhesion molecules, on PMCs increased by BCG and M. tuberculosis. These findings suggest that TLR2 plays a critical role in Mycobacteria-induced immune responses in PMCs. Keywords: Pleural mesothelial cells, Pleural effusion, Toll like receptor 2, Mycobacterium tuberculosis, BCG P-140 Monocytes are Essential for Production of Type I Interferon and Cytotoxic T Cells Response Against Mucosal Respiratory Syncytial Virus Infection Tae Hoon Kim, Heung Kyu Lee* Laboratory of Host Defenses, Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Korea Tel: 042-350-4281, E-mail: [email protected] Respiratory syncytial virus (RSV) is a leading cause of lethal respiratory infection in infants and young children worldwide. Unfortunately, no licensed vaccines and effective therapy are available. Despite its importance, current information about the cell type specific antiviral immune response against mucosal RSV infection is limited. The innate immune system senses viral invasion through pattern recognition receptors (PRRs) and evokes quick responses by producing antiviral cytokines, including type I interferon (IFN). Cell type specific function of PRRs is not well defined. Here, we report which cell types are responsible for type I IFN induction by specific TLRs or RIG-I against RSV infection. IFN-β production was dependent to MyD88 or MAVS mediated pathway in BM-DC. RSV infected BM-DM induced relatively small amounts of IFN-β, and this response was abrogated in MyD88 or MAVS deficiency. In ex-vivo study using BM cells, RSV infected plasmacytoid DC (pDC) could produce the IFN-β by TLR7-MyD88 mediated pathway. Furthermore, monocytes were also potent IFN-βproducer by MyD88 dependent manner. In early period of respiratory mucosal in vivo infection with RSV, IFN-β producing monocytes were primarily induced via the MyD88 mediated pathway. pDCs were later responder than monocytes in IFN-β production against RSV mucosal infection. Moreover, CTL response was regulated by monocytes depleted with clodronate against RSV infection. Therefore, our findings demonstrated each cell type has different antiviral mechanism against RSV infection. Particularly, in RSV infection, monocytes play a role in CTL response as well as innate response as primary type I IFN producer. Keywords: Respiratory syncytial virus, Type I interferon, Monocytes, Plasmacytoid dendritic cells The 2014 Fall Conference of the Korean Association of Immunologists 93 Poster Presentation P-141 Innate Immune Response, Infection and Inflammation Differential Control of Interleukin-6 mRNA Levels by Cellular Distribution of YB-1 Su jin Kang, Taeyun A. Lee, Eun A. Ra, Eunhye Lee, Hyun jin Choi, Sungwook Lee, Boyoun Park* Department of Systems biology, College of Life Science and Biotechnology, Yonsei University, Seoul, Korea Tel: 02-2123-8303, E-mail: [email protected] Cytokine production is essential for innate and adaptive immunity against microbial invaders and must be tightly controlled. Cytokine messenger RNA (mRNA) is in constant flux between the nucleus and the cytoplasm and in transcription, splicing, or decay; such processes must be tightly controlled. Here, we report a novel function of Ybox-binding protein 1 (YB-1) in modulating interleukin-6 (IL-6) mRNA levels in a cell type-specific manner. In lipopolysaccharide (LPS)-stimulated macrophages, YB-1 interacts with IL-6 mRNA and actively transports it to the extracellular space by YB-1- enriched vesicles, resulting in the proper maintenance of intracellular IL-6 mRNA levels. YB-1 secretion occurs in a cell type-specific manner. Whereas macrophages actively secret YB-1, dendritic cells maintain it predominantly in the cytoplasm even in response to LPS. Intracellular YB-1 has the distinct function of regulating IL-6 mRNA stability in dendritic cells. Moreover, because LPS differentially regulates the expression of histone deacetylase 6 (HDAC6) in macrophages and dendritic cells, this stimulus might control YB-1 acetylation differentially in both cell types. Taken together, these results suggest a unique feature of YB-1 in controlling intracellular IL-6 mRNA levels in a cell type-specific manner, thereby leading to functions that are dependent on the extracellular and intracellular distribution of YB-1. P-142 Evaluation of Anti-Inflammatory Effects of Ethyl Acetate Extract from Abeliophyllum distichum Leaf Kwang-Hee Kim, Joo-Hui Jeon, Yoon-Joong Kang* Department of Biomedical Science, Jungwon University, Goesan, Korea Tel: 043-830-8603, E-mail: [email protected] For many centuries, Abeliophyllum distichum has been used to treat inflammatory diseases in Goesan region as a traditional medicine. In this study, we aimed to examine anti-inflammatory activity of Abeliophyllum distichum’ leaf extract. We used ELISA and Western blotting to determine the expression level of pro-inflammatory cytokines (TNF-α, IL-6) and the phosphorylation pattern of signaling molecules of MAPK family in LPS-stimulated murine macrophage-like cell line Raw264.7 cells and human microglial cell line BV2 cells. Abeliophyllum distichum extract inhibited the production of TNF-α and IL-6. In addition, the extract suppressed the phosphorylation of ERK1/2, JNK, and p38 MAPK, and the nuclear translocation of NF-κB p65 in activated cells. These results show that Abeliophylli distichum leaf can be used as a potential therapeutic agent or functional food to relieve inflammation. Keywords: Abeliophyllum distichum, Leaf, Inflammation, Cytokine Keywords: Y-box binding protein-1 (YB-1), Secretion, IL-6 mRNA, Macrophages, Dendritic cells P-143 Vibrio vulnificus Produced-Cyclo (L-phe-pro) Inhibits Innate Immune Response Kiwan Kim, Gapryol Lee* Department of Life Science and Institute of Biological Interfaces, Sogang University, Seoul, Korea Tel: 02-705-8458, E-mail: [email protected] Cyclo (Phe-Pro) (cFP) is a secondary metabolite produced by some bacteria and fungi. Although the role of cFP in cell-to-cell communication in bacteria has been elucidated, the role of cFP in host immune response is poorly understood. In this study, we investigated the role of cFP in modulation of the innate immune responses and infection. cFP suppressed production of pro-inflammatory cytokines, nitric oxide, and reactive oxygen species in LPS-induced macrophages and bone marrow-derived macrophages. cFP inhibited IKK phosphorylation, IkBa degradation, and NF-kB translocation to the nucleus, indicating that cFP affects the NF-kB pathway. A mutant Vibrio vulnificus producing a reduced amount of cFP had lower virulence and less survival in mice compared to wild type. Therefore, cFP produced by Vibrio vulnificus actively suppresses innate immune responses of the host. Our study may shed light on the role of cFP in host-pathogen interaction and aid the development of potential therapeutic strategies for the diseases related to Vibrio vulnificus. Keywords: Cyclo (Phe-Pro), cFP, Vibrio vulnificus, NF-kB, Host-pathogen interaction 94 P-144 Ghrelin Suppresses Experimental Autoimmune Encephalomyelitis by Modulating Macrophage Polarization and Reducing CNS Inflammation Jun Ho Lee, Bonggoo Park, Hyun Seok Shin, Tae-jin Kim, Seon Ah Lim, Joanne Ng, Jiyoung Kim, Gayoung Park, Joonha Kwon, Kyung-Mi Lee* Global Research Laboratory, Department of Biochemistry and Division of Brain Korea 21 Plus program for Biomedical Science, Korea University College of Medicine, Seoul, Korea Tel: 02-920-6251, E-mail: [email protected] Thymic atrophy occurs during normal aging, and is accelerated by exposure to chronic stressors that elevate glucocorticoid levels which impair the naïve T cell output necessary to reconstitute the peripheral T-cell numbers and repertoire. The orexigenic hormone ghrelin was recently shown to attenuate age-associated thymic atrophy. Here we report that ghrelin enhances the proliferation of murine CD4+ primary T cells and a CD4+ T-cell line. Ghrelin stimulated the ERK1/2 and Akt signaling pathways, via upstream activation of phosphatidylinositol-3-kinase and protein kinase C, to enhance T-cell proliferation. Ghrelin induced the cell cycle proteins cyclin D1, E, CDK2 and phospho-Rb. Ghrelin treatment also activated these signaling pathways and stimulated thymocyte proliferation in young and older mice in vivo. Keywords: Ghrelin, EAE, Macrophage, Multiple sclerosis The 2014 Fall Conference of the Korean Association of Immunologists Innate Immune Response, Infection and Inflammation P-145 Anti-Inflammatory Effects of Ethyl Acetate Extract from Abeliophyllum distichurn Stem Jin-Wook Lee, Ji-Young Kim, Yoon Joong Kang* Department of Biomedical Science, Jungwon University, Goesan, Korea Tel: 043-830-8603, E-mail: [email protected] Abeliophyllum distichum is a medicinal plant used in regional traditional medicine to relieve pain in inflammatory processes. In this study, anti-inflammatory effects of Abeliophyllum distichum' ethyl acetate extract were examined. Furthermore, possible molecular mechanisms of the anti-inflammatory effects were dissected. The anti-inflammatory activity was investigated by inhibition of lipopolysaccharide (LPS) induced pro-inflammatory cytokine production in murine macrophage-like cell line Raw264.7 cells and human microglial cell line BV2 cells. The measurement of the induced pro-inflammatory cytokine levels were carried out by ELISA. The phosphorylation of ERK1/2, JNK, and MAPK, and the nuclear expression of nuclear factor NF-κB p65 were investigated by Western blot analysis. The extract of Abeliophyllum distichum stem significantly decreased the production of pro-inflammatory cytokines. In addition, the extract suppressed the phosphorylation of ERK1/2, JNK, and p38 MAPK, and the nuclear translocation of NF-κB p65 in activated cells. Our findings provide evidence for the popular use of Abeliophylli distichum in inflammation around Goesan region and also suggest that the stem extract has potential therapeutic benefits against several inflammatory diseases. Keywords: Abeliophyllum distichum, Inflammation, Cytokine P-147 Modulation of Dendritic Cells in the Bone Marrow during Chronic Virus Infection P-146 Poster Presentation InSAC: A Novel Subnuclear Body Involved in IL-6 RNA Processing † † Sungwook Lee , Taeyun A. Lee , Eunhye Lee, Sujin Kang, Areum Park, Boyoun Park* Department of Systems Biology, College of Life Science and Biotechnology, Yonsei University, Seoul, Korea † These authors contributed equally to this work. Tel: 02-2123-5655, E-mail: [email protected] The RNA processing of interleukin (IL)-6, an essential cytokine involved in immune regulation, must be tightly controlled during an infection. Here, we report a novel subnuclear body called the Interleukin Splicing Activating Compartment (InSAC) as a nuclear site of IL-6 mature RNA (mRNA) production. InSAC is a self-organizing structure that does not assemble as pre-formed entities in resting immune cells but instead emerges in response to LPS. InSAC contains IL-6 RNAs, small nuclear ribonucleic acids (U snRNAs), and spliceosomal small nuclear ribonucleoproteins (snRNPs), thereby facilitating IL-6 pre-mRNA processing. Importantly, disruption of InSAC formation leads to a dramatic reduction in serum IL-6 levels and its RNA processing. In addition, sustained IL-6 production in the liver and adipose tissue during obesity may be linked to LPS-independent InSAC formation. Taken together, we propose that the biogenesis of InSAC functions as a specialized compartment that is critical for efficient IL-6 RNA production during the activation of the immune response. Keywords: Splicing, InSAC, Cytokine, Obesity P-148 Anti-Inflammatory Activity of Ethyl Acetate Extract from Abeliophyllum distichum Flower Yun Hee Jeong, Sang-Jun Ha* Hwi-Gyeong Jo, Kyu-Hui Kim, Yoon-Joong Kang* Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul, Korea Tel: 02-2123-2696, E-mail: [email protected] Department of Biomedical Science, Jungwon University, Goesan, Korea Tel: 043-830-8603, E-mail: [email protected] Chronic virus infections including HIV, HBV, and HCV suppress the host immune system, resulting in the failure to the protection against persistent infections by other types of pathogens. However, mechanisms of the T cell exhaustion associated with various environments are not fully understood, especially in aspect of dendritic cells (DCs). In this study, we examined whether the immune system including DCs can be modulated in the bone marrow (BM) during chronic infection with lymphocyte choriomeningitis virus (LCMV). Since BM cells are thought to be progenitors capable of regulating immunity, we hypothesized that BM could be altered during chronic virus infection, thereby affecting T cell exhaustion. In accordance with it, we confirmed that BM was chronic virus reservoir by staining with the antibody against LCMV nucleocapsid protein (NP). When BM cells from chronically infected mice were differentiated into DCs in vitro, they expressed relatively lower levels of MHC and co-stimulatory molecules such as CD80 and CD86 but higher levels of inhibitory molecule such as PD-L1 than BM cells from naïve mice. Chronic virus infected DCs produced high levels of inflammatory cytokine IL-6, but low levels of IL-1βand TNF-α, and expressed high levels of IDO compared to the naïve BMDCs. More interestingly, CD8+ T cells primed by chronic virus infected BMDCs could not efficiently produce effector cytokines such as IFN-γ, TNF-a, and IL-2 both in vivo and in vitro, and while CD4+ T cells differentiated preferentially into Foxp3+ Treg cells in the condition of Treg induction. Taken together, it can be suggested that the progenitor cells differentiating into DCs can contribute to the T cell functional alteration during chronic virus infection. Abeliophyllum distichum is a medicinal plant used in regional traditional medicine to relieve pain in inflammatory processes. In the current study, anti-inflammatory activity of Abeliophyllum distichum’ flower extract and its possible mechanisms of action were investigated. The anti-inflammatory activity was measured by inhibition of lipopolysaccharide (LPS) induced pro-inflammatory cytokine production in murine macrophage-like cell line Raw264.7 cells and human microglial cell line BV2 cells. The measurement of the induced pro-inflammatory cytokine levels were carried out by ELISA. The phosphorylation of ERK1/2, JNK, and MAPK, and the nuclear expression of nuclear factor NF-κB p65 were investigated by Western blot analysis. Abeliophyllum distichum extract significantly decreased the production of TNF-α and IL-6. In addition, the extract suppressed the phosphorylation of ERK1/2, JNK, and p38 MAPK, and the nuclear translocation of NF-κB p65 in activated cells. The results of the present study suggest that Abeliophylli distichum flower has potential therapeutic benefits against chronic inflammation. Keywords: Abeliophyllum distichum, Flower, Inflammation, Cytokine Keywords: Chronic viral infection, Bone marrow-derived Dendritic cell, T cell The 2014 Fall Conference of the Korean Association of Immunologists 95 Poster Presentation P-149 Innate Immune Response, Infection and Inflammation Human Cytomegalovirus Inhibits TLR4 Signaling Pathway Hyun jin Choi, Areum Park, Sungwook Lee, Boyoun Park* Department of Systems Biology, College of Life Science and Biotechnology, Yonsei University, Seoul, Korea Tel: 02-2123-5655, E-mail: [email protected] The innate immune system involves innate immunological sensors, such as Toll-like receptors (TLRs), which recognizeexogenous pathogens or self-molecules as ligands in our body via infection or tissue damage to activate pro-inflammatory cytokines, such as interleukin-6 (IL-6) or interleukin-8 (IL-8), and type of interferons (IFNs) to clear harmful components. Despite such defense mechanism, human cytomegalovirus (HCMV), the largest herpesvirus that infects most of the world’s population, is able to evade immune clearance. In healthy hosts, this double-stranded DNA virus establishes life-long latent infection and stays dormant, whereas in immunocompromised hosts due to pregnancy, old age, or medications for organ transplantation, HCMV reactivates that leads to morbidity and mortality. Even in normal hosts, HCMV continuously downregulates host innate immune system for its advantage; however, knowledge of its evading strategy in host innate immunity is not well known. Here, the effect of HCMV infection during late period in TLR4 signaling was examined. This study shows HCMV infection post 48 downregulates LPS-induced interleukin-6 (IL-6) and IL-8 secretion levels. Keywords: HCMV, TLR4 P-150 Improved Effect of Statins on Metabolic Diseases by Suppressing Cardiovascular Risk Factor and Atherogenic Index in Diet-Induced Obese Mice Sungwon Lee, Kwanghee Kim, Youngjoo Lee, Junghak Kwon, Hyunseok Kong, Kyungjae Kim* College of Pharmacy, Sahmyook University, Seoul, Korea Tel: 02-3399-1601, E-mail: [email protected] Statins are a class of drugs used to lower cholesterol levels by inhibiting the enzyme HMG-CoA reductase which plays a key role in the cholesterol synthesis. It has been reported to exhibit a lipid-lowering effect on both total cholesterol and triglyceride as well as the pleiotropic effects of inflammatory disease such as atherosclerosis. The purpose of the present study was to determine the possible effect of statins on metabolic diseases including type II diabetes, hyperlipidemia and cardiovascular diseases, which are currently regarded as a main cause of death among the older generation. We examined the efficacy of two kinds of stains, Atorvastatin and rosuvastatin by applying them to DIO (Diet-induced obesity) model for metabolic disease study. The results showed that significant decrease in triglyceride, total cholesterol, LDL, and HDL levels in the animal model treated with both statins. Naturally, the both indices CRF (cardiovascular risk factor) and Al (atherogenic index) apparently diminished. Statins also showed decreased production of inflammatory cytokines such as IL-1beta and TNF-alpha as well as suppressed expression of lipid synthesis genes like SREBP1-alpha and FAS. It suggests that statins alleviate the excessive lipid accumulation and help recover the lipid balance in a good way. Taken together, we could assume the improvement in lipid metabolism can necessarily help improve cardiovascular disease such as atherosclerosis. Additionally, we also confirmed the improved effect of stain on insulin resistance by checking distinctive changes on glucose level and IPGTT. Therefore, statins are considered to have improved effect on metabolic diseases which are closely related and the fundamental cause of those diseases are consistently thought to be an obesity, statins possibly have a positive effect on various metabolic diseases by improving hyperglycemia, hyperlipidemia and hypercholesterolemia. Further research is expected to explore the mechanism behind the effects. Keywords: Statin, Metabolic Disease, Hyperlipidemia, Type II Diabetes, DIO P-151 Chronic Inflammation Affects Reactivation of Immune Cells in a Mouse Model Sun-Young Ahn, Sang-Yeon Lee, Jae-Hwan Nam* Department of Biotechnology, The Catholic University of Korea, Bucheon, Korea Tel: 02-2164-4989, E-mail: [email protected] Obesity is characterized as low-level chronic inflammation, which is thought to induce the metabolic diseases. In addition, the obese individuals may be susceptible to pathogen infection, low vaccine efficacy and worsen pathophysiology. It is previously reported that time-release of lipopolysaccharides (LPS) induced low-level chronic inflammation in a mouse model as like obese state and this inflammation contributed to increasing adiposity and aggravating glycemic control. However, it is unclear that how obesity-induced low-level chronic inflammation affect these events. In this study, we focused on the influence of LPS-induced chronic inflammation on immune cells using surrogated mouse model, which is inserted with LPS continually releasing osmotic pump. We found the reduction of macrophage activation markers in a steady-state as well as the reduction of pro-inflammatory cytokines (such as TNF-α, IL-6, IL-2, IFN-γ, and IL-4) in peritoneal macrophages and splenic T cells from mouse of chronic inflammatory model, when extra stimuli were given. Taken together, all data showed that LPS induced chronic inflammation affected the reactivation of immune cells, especially macrophages. Thus it can speculate that the dysfunction of antigen presenting cells such as macrophages might contribute to obesity-induced malfunctions. Keywords: Chronic inflammation, Peritoneal macrophage, Splenic T cells, Pro-inflammatory cytokines P-152 Protein Energy Malnutrition (PEM) Decreases Mucosal IgA Secretion Semi Rho1, Seung Young Lee1, Min Jung Kim1, Seung Hyun Shim1, Jung Hwan Kim2, Eun Jung Lee2, Myoung 2 1 Ho Jang , Jae Ouk Kim * 1 Laboratory Science Unit, International Vaccine Institute, Seoul, 2 Laboratory of Gastrointestinal Immunology, Pohang University of Science and Technology, Pohang, Korea Tel: 02-881-1318, E-mail: [email protected] Protein energy malnutrition (PEM) is caused by inadequate protein intake and this kind of malnutrition is especially common in the developing countries. PEM decreases immunity and increases infection-related morbidity. In this study, we evaluated the IgA immune responses of PEM using a mouse model. Not only nonspecific total IgA in blood but also IgA against orally administrated specific antigen (cholera toxin) was significantly increased in low protein diet (LPD) group. IgA secreting B cells in laminal propria lymphocytes (LPL) and peyer’s patches in the small intestine (SI) of LPD group were more abundant than normal protein diet (NPD) group, which indicates that B cell composition in mucosal tissue of LPD mice responds in a different way compared to the systemic immune system. In addition, LPD group showed low level of plasmablast in spleen and mesenteric lymph node, and high level of long-lived plasma cells in LPL. We next examined the form of the IgA in serum by HPLC. As a result, serum IgA in LPD group was mostly composed of polymeric form. Polymeric IgA secretion to mucosal tissue is regulated by polymeric immunoglobulin receptor (pIgR). There were decreased level of both mRNA and protein pIgR in SI of LPD group compared to NPD group. These results suggest that PEM influences B cell populations and causes a decrease of mucosal IgA secretion, which plays an important role in protection against mucosal infection. Keywords: Protein energy malnutirition, IgA, Mucosal immunity 96 The 2014 Fall Conference of the Korean Association of Immunologists Mucosal and Regional Immunity P-153 Cholera Toxin Prime Dendritic Cells to Induce Differentiation of Th17-Type CD4 T Cells through Production of Th17-Polarizing Cytokines Jung-Ok Kang, Jun Chang* Division of Pharmaceutical Sciences, Ewha Womans University, Seoul, Korea Tel: 02-3277-2549, E-mail: [email protected] Cholera toxin (CT), an exotoxin produced by Vibrio cholera, acts as mucosal adjuvant. We have previously shown that CT could skew differ+ + entiation of CD4 Th cells to IL-17-producing CD4 Th17 cells which have a crucial role in protecting extracellular pathogens like bacteria, fungi or parasites. Lung dendritic cell populations circulate between lung and draining lymph nodes regularly either in steady state or inflammation. In this study, we show that intranasally administered CT induces migration + + of migratory DC populations, CD103 DCs and CD11b DCs, to the lung + + draining lymph nodes. Comparing CD11b DCs, CD103 DCs migrate more extensively and express more MHC class II molecule and costimulatory molecules including CD40, CD80 and CD86. However even more enhanced Th17 polarization is induced in Batf3 knockout mice lacking CD103+ DCs in periphery after intranasally administrating RSV to+ gether with CT. These results demonstrate that migratory CD103 DC population in the lung tissue plays a crucial role in the differentiation of Th17 cells by intranasally administered CT. These results suggest that CT + mediate Th17 polarization through stimulating CD11b DC subset to express IL-6 and TGF-β. Keywords: Mucosal adjuvant, Cholera toxin, CD4+ Th17 cell P-154 Poster Presentation Hypoxia Increases Epithelial Permeability in Human Nasal Epithelia 1,2 1,2 Hyun Jin Min, MD, PhD , Do Yang Park , Hyo Jin 1,2 2,3 2,3 Chung , Tae Hoon Kim, PhD , Su Jin Kim , 1,2 Chang-Hoon Kim, MD, PhD * 1 Department of Otorhinolaryngology, Yonsei University College of Medicine, 2The Airway Mucus Institute, Yonsei University College of 3 Medicine, Research Center for Human Natural Defense System, Yonsei University College of Medicine, Seoul, Korea Tel: 02-2228-3621, E-mail: [email protected] The nasal mucosa is the first site to encounter pathogens, and it forms continuous barriers to various stimuli. This barrier function is very important in the innate defense mechanism. Additionally, inflammation of the nasal sinus is known to be a hypoxic condition. Here, we studied the effect of hypoxia on barrier function in normal human nasal epithelial (NHNE) cells. The expression levels of various junction complex proteins were assessed in hypoxia-stimulated NHNE cells and human nasal mucosal tissues. We performed real-time polymerase chain reaction analysis, western blotting, and immunofluorescence assays to examine differences in the mRNA and protein expression of ZO-1, a tight junction protein, and E-cadherin in NHNE cells. Moreover, we evaluated the trans-epithelial resistance (TER) of NHNE cells under hypoxic conditions to check for changes in permeability. The expression of ZO-1 and E-cadherin was measured in human nasal mucosa samples by western blotting. Hypoxia time-dependently decreased the expression of ZO-1 and E-cadherin at the gene and protein levels. In addition, hypoxia decreased the TER of NHNE cells, which indicates increased permeability. Human nasal mucosa samples, which are supposed to be hypoxic, showed significantly decreased levels of ZO-1 and E-cadherin expression compared with control. Our results demonstrate that hypoxia altered the expression of junction complex molecules and increased epithelial permeability in human nasal epithelia. This suggests that hypoxia causes barrier dysfunction. Furthermore, it may be associated with innate immune dysfunction after encountering pathogens. Keywords: Hypoxia, ZO-1, E-cadherin, Permeability P-155 Programmed Cell Death-1 (PD-1) Regulates the Resistance to DSS-Induced Colitis through Alteration of Colon Microbiota Seong Jeong Park1, Ji-Hae Kim1, Mi-Young Song2, Yunji 2 1,2 1,2 Park , Seung-Woo Lee *, Young Chul Sung 1 Department of Life Sciences, POSTECH, 2Integrative Biosciences and Biotechnology, POSTECH, Pohang, Korea Tel: 054-279-2355, E-mail: [email protected] Roles of intestinal microbiota in development of inflammatory bowel disease (IBD) have been emphasized. Recently, programmed cell death-1 (PD-1), a well-known immunosuppressive co-receptor, was found to play a critical role in regulation of bacterial composition in the gut. Here we demonstrated that PD-1 knockout (KO) mice have increased resistance to dextran sodium sulfate (DSS)-induced experimental colitis through altered bacterial communities of colon. Interestingly, PD-1 KO mice cohoused with wild-type (WT) C57BL/6 control mice manifested decreased susceptibility to the colitis. More populations of Rikenellaceae and Helicobacter muridarum were found in the cecum of WT controls compared to PD-1 KO mice. Different accumulation of inflammatory cells in the colon lamina propria (LP) of PD-1 KO mice compared to the WT mice suggested another correlation with the reduced inflammatory responses. Furthermore, altered colon microbiota of PD-1 KO mice resulted in less production of inflammatory cytokines and chemokines in colon epithelial cells. Taken together, our study indicates that PD-1 is able to improve the resistance to DSS-induced colitis by alteration of gut microbiota composition. Keywords: Inflammatory bowel disease, PD-1, DSS-induced colitis, Microbiota, Lamina propria P-156 IL-7 Produced by Intestinal Epithelial Cells in Response to Citrobacter rodentium Infection Plays a Major Role in the Innate and Adaptive Immunity against This Pathogen 1 2 1 Wei Zhang , Qing Yu , Jun-O Jin * 1 Shanghai Public Health Clinical Center, shanghai medical college, Fudan 2 University, Shanghai, China, Department of Immunology and infectios diseases, The Forsyth Institute, Cambridge, MA, USA Tel: 051-624-9744, E-mail: [email protected] IL-7 plays a pivotal pathogenic role in inflammatory bowel diseases, but the role of IL-7 in intestinal bacterial infection remains unclear. Here we characterized the previously unexplored role of IL-7 in the innate and adaptive immune response to attaching-and-effacing bacterium C. rodentium infection. We examined the role of endogenous IL-7, which produced by intestinal epithelial cells in response to C. rodentium, in C. rodentium-infection. Treatment with anti-IL-7Rα Ab during C. rodentium infection resulted in higher bacterial burden, enhanced intestinal damage, and greater weight loss and mortality compared to control IgG treatment. The impaired bacteria clearance upon IL-7Rα blockade was associated with a significant decrease in Th1 and Th17 responses, CXCR3 ligand expression and macrophages accumulation and activation in the colon. Moreover, C. rodentium-induced expansion and activation of intestinal CD4+ LTi cells was completed abrogated by IL-7Rα blockade. In addition, IL-7Rα blockade during C. rodentium infection promoted recruitment and activation of neutrophils in colon, which may potentially enhance tissue damage. These data suggest that IL-7 is produced by intestinal epithelial cells in response to C. rodentium-infection and promotes the recruitment and function of innate and adaptive immune cells for the clearance of these bacteria and protection of intestinal tissues. IL-7 plays a critical role in the protective immunity against intestinal attaching-and-effacing bacterium infection. Keywords: IL-7, C. rodentium, effector T cell, Macrophage, CD4+ LTi cell The 2014 Fall Conference of the Korean Association of Immunologists 97 Poster Presentation P-157 Mucosal and Regional Immunity Bacterial β-(1,3)-Glucan Prevents DSS-Induced IBD by Restoring the Reduced Population of Regulatory T Cells P-158 Short-Term Fasting Protects Mice Infected with + Listeria monocytogenes by Induction of Foxp3 Tregs through Migratory CD103+ DC Kon-Young Ji1, Mi-Kyoung Kim2, In-Young Lee2, Ha-Rim Choi3, Hyung-Sik Kang1* Kyungmin Lee1, Girak Kim1, Seung-Hyun Han3, Cheol-Heui Yun1,2* 1 1 Bacterial β-(1,3)-glucan has more advantages in terms of cost, yield and efficiency than that derived from mushrooms, plants, yeasts and fungi. We have previously developed a novel and high-yield β-(1,3)-glucan produced by Agrobacterium sp. R259. This study aimed to elucidate the functional mechanism and therapeutic efficacy of bacterial β-(1,3)-glucan in dextran sulfate sodium (DSS)-induced inflammatory bowel disease (IBD). Mice were orally pretreated with bacterial β-(1,3)-glucan at daily doses of 2.5 or 5 mg/kg for 2 weeks. After 6 days of DSS treatment, clinical assessment of IBD severity and expression of pro-inflammatory cytokines were evaluated. In vivo cell proliferation was examined by immunohistochemistry using Ki-67 and ER-TR7 antibodies. The frequency of regulatory T cells (Tregs) was analyzed by flow cytometry. Natural killer (NK) activity and IgA level were evaluated using NK cytotoxicity assay and ELISA. The deterioration of body weight gain, colonic architecture, disease score and histological score was recovered in DSS-induced IBD mice when pretreated with bacterial β-(1,3)-glucan. The recruitment of macrophages and the gene expression of proinflammatory cytokines, such as IL-1β, IL-6 and IL-17A/F, were markedly decreased in the colon of β-(1,3)-glucan-pretreated mice. β-(1,3)-Glucan induced the recovery of Tregs in terms of their frequency in DSS-induced IBD mice. Intriguingly, β-(1,3)-glucan reversed the functional defects of NK cells and excessive IgA production in DSS-induced IBD mice. We conclude that bacterial β-(1,3)-glucan prevented the progression of DSS-induced IBD by recovering the reduction of Tregs, functional defect of NK cells and excessive IgA production. Fasting is literally the prohibition of food uptake causing nutritional depletion. Even though gastrointestinal tract is the largest organ producing and consuming a great amount of energy and the first organ to be directly affected after the fasting, very little is known about how fasting influences intestinal immune cells. Innate immune cells in gastrointestinal tract play a critical role as an initial sensor of antigens and inducer of proper immune response. In the present study, we focused on the changes of intestinal immune cells in mice upon 24 hrs short-term fasting and how the fasting influences the protection against L. monocytogenes infection. The results showed that the mice with short-term fasting increased a survival rate compared to control (ad libitum) mice infected with L. monocytogenes through intragastric route. No differences on L. monocytogenes colony forming unit (cfu) between the two groups were found in spleen, liver and mesenteric lymph nodes (mLN) at 3, 9, 24 and 48 hrs post infection. It is intriguing that the number of CD103+CD11b− DCs in both small intestinal lamina propria and mLN were significantly increased after either fasting or fasting followed by infection. Furthermore, the expression of CCR7 was up-regulated on CD103+CD11b− DCs in mice had been short-term fasting and infected with L. monocytogenes. In addition, the CD103+CD11b− + DCs owing to fasting induced more Foxp3 Tregs in mLN than DCs from control mouse. Taken together, these results suggested short-term fasting can induce the tolerogenic condition in intestine through increased CD103+ CD11b− DCs which might play an important role in survival of mice against L. monocytogenes infection. Keywords: Bacterial β-(1,3)-glucan, Inflammatory bowel disease, Dextran sulfate sodium (DSS), Regulatory T cell, Natural killer cell, Immunoglobulin A Keywords: Fasting, Regulatory T cells, CD103 dendritic cells, Immune tolerance, Listeria monocytogenes School of Biological Sciences and Technology, Chonnam National University, Gwangju, 2Naturence Co., Ltd. Tanchun Industrial Complex, 3 Gongju, Department of nursing science, Nambu University, Gwangju, Korea Tel: 062-530-2195, E-mail: [email protected] P-159 Microbial Metabolite Regulates Mucosal Immune Homeostasis through Modulation of Innate Lymphoid Cells 1,2 2 1,2 Sae-Hae Kim , Ha-Yan Lee , Yong-Suk Jang * 1 Department of Molecular Biology and the Institute for Molecular Biology and Genetics, 2Department of Bioactive Material Sciences and Research Center of Bioactive Materials, Chonbuk National University, Jeonju, Korea Tel: 063-270-3343, E-mail: [email protected] Innate lymphoid cells (ILCs) were recently characterized as a group of immune cells closely located at barrier surfaces of mucosal lymphoid organs. ILCs are known to be associated with several responses including lymphoid organogenesis, tissue remodeling, antimicrobial immunity, and inflammation through cytokine-mediated regulation. ILC subsets can be categorized into two separate lineages such as cytotoxic ILCs and cytokine-producing helper-like ILCs (i.e., ILC1s, ILC2s, ILC3s). In mucosal immunity, ILC3s play an essential role for early protection against enteric pathogens, although regulation of the ILCs by mucosal environment is not clearly understood yet. In this study, we suggest butyrate, one of the metabolites produced by commensal microbes, as an ILC modulator in Peyer’s patch based on the observation that butyrate treatment is closely associated with protection against the bacterial infection and DSS colitis. In order to understand the underlying mechanism on modulation of ILCs exerted by butyrate, we monitored the expression pattern of transcription factors and cytokines in Peyer’s patch ILCs after oral administration of sodium butyrate into SPF, nude, and antibiotics-treated mice. In addition, in order to understand the function of butyrate-induced Peyer’s patch ILCs, we analyzed the immune response using DSS colitis model in SPF and nude mice. Collectively, the results suggest that regulation of ILCs by butyrate was closely associated with the maintenance of mucosal homeostasis in gastrointestinal tract. This study was supported by the NRF, 2013R1A2A2A01014459. Dr. S.-H. Kim and Ms. H.-Y Lee were supported by the BK21 Plus program in the Department of Bioactive Material Sciences. Keywords: Butyrate, Innate lymphoid cells, Mucosal homeostasis 98 Department of Agricultural Biotechnology and Research Institute for Agriculture and Life Sciences, Seoul National University, 2Biomodulation major and Center for Food and Biocenvergence, Seoul National University, 3Department of Oral Microbiology and Immunology, DRI and BK21 Program, School of Dentistry, Seoul National University, Seoul, Korea Tel: 02-880-4802, E-mail: [email protected] + P-160 The Cathelicidin LL-37 is Capable of Playing as a Mucosal Adjuvant in Oral Vaccine Model Sae-Hae Kim1,2, Ha-Yan Lee2, Yong-Suk Jang1,2* 1 Department of Molecular Biology and the Institute for Molecular Biology and Genetics, 2Department of Bioactive Material Sciences and Research Center of Bioactive Materials, Chonbuk National University, Jeonju, Korea Tel: 063-270-3343, E-mail: [email protected] Tight regulation of the immune response interconnecting innate and adaptive immunity is critical in host defense against pathogen infections. Consequently, it is assumable that antimicrobial peptide could act as a mediator to link and regulate both stages of immune responses. Human cathelicidin antimicrobial peptide LL-37 was previously identified to have adjuvant activity in parenteral vaccination procedure, although similar activity is not analyzed in mucosal immune system. In this study, we characterized adjuvant activity of LL-37 in oral immunization. Oral mucosal immunization of LL-37-conjugated EGFP effectively evoked the antigen-specific systemic and mucosal immune responses. In addition, when the LL-37 was applied to pathogenic antigen, an envelope domain III of dengue virus type-2, antibody responses with high neutralization activity was induced. Further analysis in Peyer’s patch lymphocytes after oral administration of LL-37-conjugated antigen showed the increase in number of germinal centers in Peyer’s patch and mesenteric lymph node by chemotactic effect of LL-37. Oral priming with LL-37-conjugated antigen induced Th1- and Th17-skewed immune responses. More interestingly, we found the expression of formyl peptide receptor-2, one of the receptors for LL-37, in M cells and effective delivery of LL-37 conjugated antigen into the receptor-expressing cells. Collectively, we concluded that LL-37 can play a role as mucosal adjuvant through enhanced antigen delivery and antigen-specific immune induction. This study was supported by the NRF, 2013R1A2A2A01014459. Dr. S.-H. Kim and Ms. H.-Y Lee were supported by the BK21 Plus program in the Department of Bioactive Material Sciences. Keywords: Mucosal vaccine, Mucosal adjuvant, Cathelicidin LL-37 The 2014 Fall Conference of the Korean Association of Immunologists Mucosal and Regional Immunity P-161 Strong Association between SNPs in the IL6 and IL6R Genes and Korean Dry Eye Disease Patients P-162 Poster Presentation Tear Expression Profiling of Cytokine, Chemokine and Soluble Receptor in Korean Keratoconus Patients YoungSik Yoo1, HeeJung Ju1, Kyung-Sun Na1,2,3, Jee-Won Mok1, Choun-Ki Joo1,2,3* Young-Sik Yoo1, Kyung-Sun Na1,2,3, Jee-Won Mok1, Choun-Ki Joo1,2,3, Yong-Soo Byun1* 1 1 Catholic Institutes of Visual Science, The Catholic University of Korea, Department of Ophthalmology & Visual Science, College of Medicine, The 3 Catholic University of Korea, Seoul St. Mary’s Hospital Eye Institute (SSEI) Tel: 02-2258-6906, E-mail: [email protected] Catholic Institutes of Visual Science, The Catholic University of Korea, Department of Ophthalmology & Visual Science, College of Medicine, The 3 Catholic University of Korea, Seoul St. Mary’s hospital Eye Institute (SSEI) Tel: 02-2258-6906, E-mail: [email protected] To investigate the possibility of IL6 and IL6R as major susceptibility genes for Korean Dry eye disease patients. Genomic DNA was extracted from blood samples of unrelated dry eye disease patients. PCR and direct sequencing were used to screen genetic variations. One hundred fifty control individuals without corneal disease were selected from the general population. We investigated 8 SNPs for IL6 and 5 SNPs of IL6R. Among them, rs1800796 (-572 c>g, OR=2.571) in 5’UTR of IL6 and rs4845617 (EX1 +230 g>a, OR=1.769) and rs2228145 (D358A , OR=1.119) in exon of IL6R were significantly different between patients and control groups. The genotype distributions of all polymorphisms of IL6 and IL6R among the control subjects and the affected individuals were in Hardy-Weinberg equilibrium. The present study showed that the genetic variants in IL6 and IL6R, in the rs1800796 (-572 c>g) of IL6 and rs4845617 (EX1 +230 g>a) and rs2228145 (D358A) of IL6R, are associated with a higher risk of dry eye disease patients. It is suggested that genetic variations of IL6 and IL6R may act as a potential susceptibility variants in Korean Dry eye disease. To determine whether the expression levels of cytokines, chemokines and soluble receptor in tear of keratoconus patients contribute to the pathogenesis of keratoconus. The patients with keratoconus were diagnosed based on the following criteria: Munson sign, protrusion, Vogt’s striae, corneal thickness, scarring, Fleishcher ring, photokeratoscopy signs and refractive errors and medical histories, e.g., eye rubbing and atopy. Tears were collected from 28 keratoconus patients (56 eyes) and 30 healthy subjects (60 eyes) by a polyurethane minisponges. Control subjects with no history of ocular disease were also enrolled. The concentrations of cytokines/chemokines were analyzed by Luminex 200 using Human cytokine/chemokine 65 molecules and Human soluble cytokine receptor (14 molecules). The Median Fluorescent Intensity (MFI) was used to obtain the calculating cytokines, chemokines and soluble receptor concentrations in tears. Of the 79 cytokines, chemokines and soluble receptors, we detected 16 molecules that demonstrated significant differences in tears from Keratoconus patients. In cytokines, G-CSF (p<0.001), IL1-ra (p<0.05), VEGF (p<0.05), IL-16 (p<0.05) and IL-20 (p<0.05) were significantly increased in Keratoconus patients compared to control subjects. MDC (p<0.05) and GRO (p<0.05) of chemokines and sIL-1RI (p<0.05) and sIL4R (p<0.001) of soluble receptors were increased in keratoconus patients. Whereas IL4 and IFN gamma (all p<0.001) and FGF2 ( p<0.05) of cytokines, MIP1a and MIP1b (all p<0.001) of chemokines, sCD30 and sIL6R (all p<0.05) were significantly decreased in keratoconus patients. In tears of keratoconus patients, nine molecules were elevated, whereas 7 molecules were decreased. It is suggested that different levels of inflammatory regulated molecules may all play an important role in the pathogenesis of keratoconus. 2 Keywords: Dry eye, IL6, IL6R, Association 2 Keywords: Keratoconus, Tear, Cytokine, Chemokine, Soluble receptor P-163 Vitamin A Deficiency Alters Ocular Surface Barrier Soo Jung Han, Kyoung Yul Seo* Department of Ophthalmology, Eye and Ear Hospital, Severance Hospital, Institute of Vision Research, Yonsei University College of Medicine, Seoul, Korea Tel: 02-2228-0748, E-mail: [email protected] Vitamin A has been shown to regulate the growth and differentiation of epithelial cells in many tissues, and it has both positive and negative regulatory functions in the immune system. To study the effects of depletion of retinoic acid on ocular surface and lacrimal immune system, we generated vitamin A-deficient (VAD) mice by continuous feeding of a VAD diet beginning in gestation. We found that muc5AC concentration was significantly reduced in tear of VAD mice without alteration of other antimicrobial peptide. Accordingly, total amount of bacteria was significantly increased in the ocular surface of vitamin A-deficient (VAD) diet -fed mice. Furthermore, in the lacrimal gland of VAD diet-fed mice, IFN-g-secreting CD4+ T cells had significantly been increased than those of the control diet-fed mice. Taken together, these data indicate that vitamin A deficiency interferes with the integrity of the ocular mucosal barrier. Keywords: Vitamin A deficiency, Ocular surface, Muc5AC, Lacrimal gland P-164 Lactoferrin and Retinoic acid Synergize to Enhance Gut IgA Response Bo-Ra Jin1, Jeong-Min Lee1, Seong-Ho Kang1, Young-Saeng Jang1, Goo-Young Seo1, Woan-Sub Kim2, 1 Pyeung-Hyeun Kim * 1 Department of Molecular Bioscience, College of Biomedical Science, 2 Kangwon National University, Department of Animal Life and Environmental Science, Hankyong National University, Korea Tel: 033-250-8546, E-mail: [email protected] Both lactoferrin (LF) and retinoic acid (RA) are abundant in the colostrum and milk. Recently, we observed that TGF-β1 in cooperation with RA increases IgA isotype switching and expression of gut homing molecules (CCR9, α4β7) in mouse. In the present study, we investigated the effect of LF and RA in combination on IgA B cell differentiation. LF and RA in combination increased IgA production as well as IgA germ line transcription. These two molecules also stimulated splenic B cells to express gut-homing molecules and differentiate into plasma cells. Further, such effects of LF and RA were also observed for peritoneal B1 cells. These results suggest that LF in cooperation with RA contributes to overall gut IgA Ab response. Keywords: Retinoic acid, Lactoferrin, IgA, Gut homing molecule The 2014 Fall Conference of the Korean Association of Immunologists 99 Poster Presentation P-165 Mucosal and Regional Immunity Commensal Microbiota and Food Antigens are Important Factors in the Regulation of IgE Production in Mice Sung-Wook Hong1,2, Kwangsoon Kim1,2, Charles D. Surh1,2,3* 1 Academy of Immunology and Microbiology (AIM), Institute for Basic Science (IBS), 2Department of Integrative Biosciences and Biotechnology 3 (IBB), Pohang University of Science and Technology, Pohang, Korea, La Jolla Institute for Allergy and Immunology, USA Tel: 054-279-8721, E-mail: [email protected] IgE is one of key modulators in the pathogenesis of allergy by inducing FcR-mediated activation of mast cells and basophils. Production of IgE is regulated by commensal microbiota, as it is normally undetectable in conventional mice, but highly elevated in germ-free mice shortly after weaning into solid food. The mechanism behind IgE production in germ-free mice is poorly understood, but could be a consequence of dysregulated immune responses to Ags in the food. To address this idea, germ-free mouse pups were weaned into an antigen-free elemental diet comprised of essential amino acids, vitamins and minerals. Strikingly, neither these mice nor their offsprings, designated as antigen-free mice, possessed detectable levels of serum IgE. Moreover, feeding adult germ-free mice with antigen-free diet reduced the serum IgE level, indicating continued presence of food is required to sustain high amount of IgE production. As expected, antigen-free mice weaned into normal solid food diet produced high levels of IgE. To determine the age-dependency of commensal microbiota in IgE production, germ-free mice were conventionalized at various ages, starting from 4 wks. Colonization of commensal microbiota at 4 wks of age prevented high levels of serum IgE, but not in older mice. These results suggest that food Ags by default induce a strong IgE response that is suppressed by the commensal microbiota. Keywords: IgE, Germ-free, Microbiota, Food antigens P-166 Tonsil-Derived Mesenchymal Stromal Cells Acquired a Follicular Dendritic Cell Phenotype Under Toll-Like Receptor 3 (TLR3) and TLR4 Stimuli and Recruited CXCR2 Expressing Cells 1 1 1 Jung-Hwa Ryu , Minhwa Park , Bo-Kyung Kim , 2 1 Kyung-Ha Ryu , So-Youn Woo * 1 Department of Microbiology, and 2Pediatrics, School of Medicine, Ewha Womans University, Seoul, Korea Tel: 02-2650-5737, E-mail: [email protected] Previously, we have shown that mesenchymal stromal cells (MSCs) inhibit graft-versus-host disease (GVHD) of mice and macrophage inflammatory protein-2/CXCR2 blockade attenuates acute GVHD while preserving graft-versus-leukemia activity. Although MSCs have known for regulatory effect on immune or inflammatory responses, migration onto the specific organ site and local interactions were not much studied. We hypothesized that migration into the specific organs or local areas after injection of MSCs and exert immune modulatory effects might be involved in Toll-like receptor-mediated stimuli. We used the tonsil-derived mesenchymal stromal cells (T-MSCs), which had the potential to differentiate into mesenchymal origin, and analyzed the expression profile of TLRs on T-MSCs and stimulated T-MSCs with each of TLR agonists. We found that in case of T-MSCs, TLR3 stimuli induced the CCR6 expression after 24 hrs. Further, Cytokine Array Analysis showed that epithelial-derived neutrophil attractant-78 (ENA78/CXCL5), granulocyte chemotactic protein-2 (GCP2/CXCL6), growth-related gene product α (Groα /CXCL1), IL-8/CXCL8, and interferon-inducible protein-10 (IP-10/CXCL10) were increased. In addition, CD23, CD35 and CD54 expression were increased after TLR3 and TLR4 stimulation. Therefore, it suggested that TLR3 or TLR4 stimulation on T-MSCs led to differentiate into follicular dendritic cells and secreted chemokines, possibly recruiting CXCR2-expressing immune cells. This work was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2012M3A9C6049823). Keywords: Follicular dendritic cells, Toll-like receptor, CXCR2, Mesenchymal stromal cells P-167 IL-10 Producing CD8+ T Cells and Plasmacytoid Dendritic Cells Have Compensatory Role to Maintain Gut Immune Regulation P-168 Visualization of Inoculated Eyedrop Draining Passages and Identification of Eyedrop Draining Lymphnodes and Antigen Presenting Cells in Eyedrop Vaccination 1† 2† 2 Young-In Kim , Bo-Ra Lee , Hyun-Jeong Ko , 1 Sun-Young Chang * 1,4 2 3 Eun-Do Kim , Soo Jung Han , Kyoung Sub Choi , 2 2 Sangchul Yoon , Kyoung Yul Seo * 1 College of Pharmacy, Ajou University, Suwon, 2College of Pharmacy, Kangwon National University, Chuncheon, Korea † Equally contributed. Tel: 031-219-3454, E-mail: [email protected] 1 The Graduate School of Yonsei University, 2Department of Ophthalmology, Eye and Ear Hospital, Severance Hospital, Institute of Vision Research, 3 Yonsei University College of Medicine, Seoul, Department of Ophthalmology, National Health Insurance Service Ilsan Hospital, Goyang, 4Brain Korea 21 Project for Medical Science, Yonsei University, Seoul, Korea Tel: 02-2228-3581, E-mail: [email protected] + CD4 regulatory T cells have been investigated to be a critical regulator to maintain gut immune suppressive environments since Foxp3 gene was reported as transcription factor for CD4+ Treg differentiation. To exert sup+ pressive function of CD4 Tregs, CCR7 is important which mediates the migration process of them from mucosal periphery into draining lymph node. Here, we hypothesized whether CCR7 deficiency fail to establish immune suppression in the DSS-induced colitis. Unexpectedly, intestinal inflammation was not exacerbated in absence of CCR7. IL-10, a representative suppressive cytokine was enhanced in the CD8+ T cells in the + lamina propria of CCR7 KO colon. CD8 T cells isolated from lamina propria of CCR7KO colon could reduce the activation of CD4+ T cell. Plasmacytoid dendritic cells which express PDCA-A were also slightly increased under intestinal inflammation in absence of CCR7. These suggested that IL-10 producing CD8+ T cells and plasmacytoid dendritic cells have compensatory role to maintain gut immune regulation in functional + defect of CD4 Treg. Keywords: Colon, Inflammation, CCR7, CD8+ T cell, Plasmacytoid dendritic cell In our previous study, the level of immune induction and the efficacy of eyedropand IN vaccine has been compared, and IN vaccination showed significantly stronger level of immune induction. However, the eyedrop vaccination activates both tear-associated lymphoid tissues (TALT) and nasal-associated lymphoid tissues (NALT) which are used for the induction of immunity and antigen uptake in intranasal immunization. Moreover, there were only T cell proliferation assay for the detection of draining lymphnodes of eyedrop vaccination. Therefore, the clarification of the draining passageway of eyedrop vaccination and the comparison of the draining lymphnodes between those two different entry sites are needed. In here, we firstly visualized the route of drainage of the eyedrop vaccine and showed that the draining lymphnodes for eyedrop vaccination are different from those of intranasal vaccination in mice. The visualizing materials administered by eyedrop drained out through tear duct and also via nasal cavity. To contrast the draining lymphnodes between eyedrop and intranasal vaccination, FITC solution and FITC labeled-beads were used. After the FITC solution was injected by eyedrop, superficial parotid lymphnodes (SPLN), mandibular lymphnodes (MdLN), inguinal lymphnodes (InLN) and spleen (SPL) were all FITC positive. However, 2 days after FITC-beads inoculation by eyedrop, superficial parotid lymphnodes (SPLN) were bead positive only in the presence of poly(I:C) and little mandibular lymphnodes (MdLN) were bead positive. In contrast, intranasally administered beads were detected in both SPLN and MdLN. Among FITC-beads positive cells in SPLN, they were CD11c+CD86+ or F4/80+. These results suggest that inoculated eyedrop vaccine antigens are captured by CD11c+ DCs and possibly participate in the presentation of the antigens to T cells in SPLN, rarely in MdLN, which a pattern is distinguished from that of IN vaccination. Keywords: Eyedrop vaccination, Lymphnode 100 The 2014 Fall Conference of the Korean Association of Immunologists Tumor and Transplantation Immunology P-169 Regulation of Nod2-Linked Pro-inflammatory Signals by Mucosal Ribotoxic Stress 1 2 1 Seong-Hwan Park , Dongwook Kim , Yuseok Moon * 1 Laboratory of Mucosal Exposome and Biomodulation, Department of Microbiology and Immunology, Immunoregulatory Therapeutics Group in Brain Busan 21 Project, Pusan National University School of Medicine, 2 Yangsan, National Institute of Animal Science, RDA, Suwon, Korea Tel: 051-510-8094, E-mail: [email protected] In response to excessive nucleotide-binding oligomerization domain−containing protein 2 (Nod2) stimulation caused by mucosal bacterial components, gut epithelia need to activate regulatory machinery to maintain epithelial homeostasis. Activating transcription factor 3 (ATF3) is a representative regulator in the negative feedback loop that modulates TLR-associated inflammatory responses. In the current study, the regulatory effects of ribotoxic stress-induced ATF3 on Nod2-stimulated pro-inflammatory signals were assessed. Ribotoxic stress caused persistent ATF3 expression that in turn suppressed pro-inflammatory chemokine production facilitated by Nod2. Decreased chemokine production was due to attenuation of Nod2-activated NF-kB and early growth response protein 1 (EGR-1) signals by ATF3. However, the underlying molecular mechanisms involve two convergent regulatory pathways. Although ATF3 induced by ribotoxic stress regulated Nod2-induced EGR-1 expression epigenetically through the recruitment of histone deacetylase 1, NF-kB regulation was associated with posttranscriptional regulation by ATF3 rather than epigenetic modification. ATF3 induced by ribotoxic stress led to the destabilization of p65 mRNA caused by nuclear entrapment of transcript-stabilizing human Ag R protein via direct interaction with ATF3. These findings demonstrate that ribotoxic stress-induced ATF3 is a critical regulator in the convergent pathways between EGR-1 and NF-kB, which contributes to the suppression of Nod2- activated pro-inflammatory gene expression. This work was carried out with the support of “Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ008405032014)” Rural Development Administration, Republic of Korea. Keywords: Nod2, Ribotoxic stress, ATF3, HuR, Pro-inflammatory signals P-171 Functional Properties of THP-1 Cells-Derived M1 and M2 Macrophages Su Jung Oh1, Soo Kyung Jeong1, Kwangmo Yang1, Chang Geun Lee1, You Soo Park1, Joong Sun Kim1, Sung Dae 1 2 2 1 Kim , So Young Park , Min Ho Jeong *, Wol Soon Jo * 1 Department of Research Center, Dong Nam Institute of Radiological and 2 Medical Sciences, Department of Microbiology, Dong-A University College of Medicine, Busan, Korea Tel: 051-720-5013, E-mail: [email protected] Mononuclear phagocytes are versatile cells that can express different functional programs in response to microenvironmental signals. Fully polarized M1 and M2 macrophages are the extremes of a continuum of functional states. Macrophages that infiltrate tumor tissues are driven by tumor-derived and T cell-derived cytokines to acquire a polarized M2 phenotype. The aim of the present study was to design an accessible and convenient in vitro model to study macrophage morphology and functions in relation to the tumor microenvironment. The differentiation of THP1 cells in response to PMA and polarization of THP1-derived macrophages in response to IFN-γ or IL-4/IL-13 were examined comparing to M1and M2-polarized macrophages (or Tumor associated macrophage, TAM). When treated with PMA and cytokines, differentiated M1 or M2 macrophages showed dramatically differences in cytoplasmic volume, the intensity of mitochondrial and lysosomal staining, surface expression (CD 206, CD204, CD36, CD80, MHC class II), phagocytic capacity and PLT aggregation. The differentiated M1 or M2 macrophages also induced different pattern in production of inflammatory cytokine (IL-1β, TNF-αand IL-6) and immunosuppressive cytokines (IL-10 and TGF-β). In the role of macrophages in tumor invasion, MDA-MB-231 cells co-cultured with THP-1 cells derived by PMA only and M-2 polarized cells showed an increased number of cells invading through 8 μm pores on the lower side of filter compared to M-1 polarized cells. In addition, THP-1 cells derived by PMA only and M-2 polarized cells significantly induced the expression of MMP-9 of MDA-MB-231 cells compared to M-1 polarized cells. Taken together, morphological and functional analysis illustrated that while PMA stimulation induced macrophage differentiation with M2-like phenotypes, important differences existed in comparison to TAM. A protocol in which THP1 cells were activated with PMA then exposed to IL-4/IL-13 more closely resembled the phenotype of TAM. P-170 Poster Presentation Interferon Gamma Induced by Resveratrol Analog, HS-1793, Reverses the Properties of Tumor Associated Macrophages Soo Kyung Jeong1, Kwangmo Yang1, You Soo Park1, Chang Geun Lee1, Joong Sun Kim1, Sung Dae Kim1, Su Jung Oh1, So Young Park2, Min Ho Jeong2*, Wol Soon Jo1* 1 Department of Research Center, Dong Nam Institute of Radiological and 2 Medical Sciences, Department of Microbiology, Dong-A University College of Medicine, Busan, Korea Tel: 051-720-5013, E-mail: [email protected] Macrophages are capable of both inhibiting and promoting the growth and spread of cancers, depending on their activation state. Tumor-associated macrophages (TAM) are a kind of alternatively activated M2 macrophage, which may contribute to tumor progression. Following our previous study to evaluate the anti-tumor effect of a synthetic resveratrol analog HS-1793, the current study demonstrated that HS-1793 treatment significantly increased IFN-γ secreting cells in splenocytes and decreased CD206+ macrophage infiltration compared to CD68+ cells in the tumor site with a higher expression of IFN-γ. As these results suggested that IFN-γ increased locally at the tumor sites could modulate the status of TAM, we designed an in vitro model to study macrophage morphology and functions in relation to the tumor microenvironment. Human monocytic cell line THP-1 cells stimulated with phorbol-12-myristate-13-acetate (PMA) differentiated to macrohigh high phageshighwith M2-like phenotypes. TAM-like properties of CD206 , CD204 , high low ow high high IL-10 , TGF-β , IL-6 , IL-12 , VEGF , and MMP-9 and promotion of tumor cell invasion were more pronounced inM-2-polarized THP-1macrophages generated by differentiating THP-1 cells with PMA and subsequently polarizing them with Th2 cytokines (IL-4/IL-13). Upon IFN-γ exposure, THP-1derived TAM changed their phenotypes to the M-1-like morphology and intracellular granular pattern with an expression of an increased level of proinflammatory and immunostimulatory cytokines and a reduced level of immunosuppressive and tumor progressive mediators. These results explain the underlying mechanism of the anti-tumor activity of HS-1793. The elevated level of IFN-γ production after HS-1793 treatment evoked reprogramming of M-2 phenotype TAM, which efficiently countered the immunosuppressive and tumor progressive influences of TAM. Keywords: HS-1793, IFN-γ, Tumor-associated macrophages, M-2 polarized macrophages, THP-1-derived macrophages, Tumor progression P-172 Comparative Immunological Study of Lamellar and Penetrating Keratoplasty in Murine Model Hyun Soo Lee, Min-Young Choi, Choun-Ki Joo* Department of Ophthalmology and Catholic Institute for Visual Science, Seoul St. Mary’s Hospital, Seoul, Korea Tel: 02-2258-1173, E-mail: [email protected] Background and Purpose: Lamellar keratoplasty (LK) has been considered an acceptable alternative surgery for penetrating keratoplasty (PKP), because LK has the advantage of lower risk of graft rejection and intraocular complications, according to recent clinical outcomes. But the relative immunological mechanism of graft rejections for the two procedures is uncertain. To understand the immunological mechanisms of lamellar keratoplasty (LK) and penetrating keratoplasty (PKP) in murine model. Methods: PKP or LK was performed C57BL/6 donor grafts and BALB/c recipients and graft opacity was assessed. We evaluated immunologic responses with delayed-type hypersensitivity (DTH) assays, real-time PCR, and immunohistochemistry. Results: LK mice showed less graft rejection than PK mice and LK led to less DTH response and IFN-γ secretion in vitro recall assay of T cells from drainage lymph nodes, as compared to PK (acceptance; acc). There was no difference of mRNA expressions of proinflammatory cytokines in the corneas early (2wk), but those of PKacc mice were significantly increased later (4wk). In addition, LK showed less angiogenesis and lymphangiogenesis in grafted cornea than PK, and LK led to decreased number of mature MHC-IIhighCD11c+ cells in the draining LNs and less infiltration of CD3+ T cell into the grafted corneas of transplanted mice. Conclusion: LK presents less graft rejection than PK through lower angiogenesis and lymphangiogenesis, which could make pathologic T cells and APC less migration into drainage LNs and grafted corneas, respectively. Keywords: Cornea, Tranplantation, T cells Keywords: Mononuclear phagocytes, THP-1 cells, M1 and M2 macrophages, Tumor associated macrophage The 2014 Fall Conference of the Korean Association of Immunologists 101 Poster Presentation P-173 Tumor and Transplantation Immunology Reduction in Renal Ischemia-Reperfusion Injury in Mice by a Phosphoinositide 3-Kinase p110gamma-Specific Inhibitor 1 1 1 Nayoung Kim *, Dong-Cheol Woo , Youyol Song , Seokmann Hong2, Yong Mee Cho3, Duck-Jong Han4 1 Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, 2Deptment of Bioscience and Biotechnology, Institute 3 4 of Bioscience, Sejong University, Department of Pathology, and Department of Surgery, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea Tel: 02-3010-2624, E-mail: [email protected] Background: Although renal ischemia-reperfusion injury (IRI) can cause delayed graft function, a targeted therapy is not yet available. Because phosphoinositide 3-kinase (PI3K) p110γand p110δplay important roles in immune cell migration and function, we investigated the effects of PI3K p110γ- and p110δ-specific inhibitors in a murine renal IRI model. Methods: Renal function was assessed by serum creatine and hematoxylin and eosin staining. Immune cell migration was assessed by flow cytometry and using an in vitro cell migration assay. Magnetic resonance imaging was performed to investigate the cellularity and microstructure of the kidneys. To understand the mechanism of the therapeutic effect of p110γ-specific inhibitor, we performed a multiplex cytokine/chemokine assay and western blotting. Results: PI3K p110γ-specific inhibitor, but not p110δ-specific inhibitor, significantly reduced serum creatine levels and acute tubular necrosis. This was accompanied by reduced infiltration of B and T cells and reduced expression of a CXCR3 ligand, suggesting that p110γplays an important role in T and B cell migration toward injured kidneys. An in vitro cell migration assay revealed for the first time that the migration of B cells to injured kidney cells requires p110γ. Conclusions: p110γ-specific inhibitor ameliorates renal IRI by reducing necrosis and immune cell migration. This inhibitor may have the potential to reduce renal graft failure caused by renal IRI. Keywords: Renal ischemia-reperfusion injury, PI3K p110gamma-specific inhibitor, Lymphocyte migration P-175 P-174 1,2 2 1 Soseul Kim , Jeong won Hong , Woon-Dong Cho , Yoo 1,2 2 1 Ri Moon , Sang Soon Yoon , Min-Young Kim , Kwon Pyo Hong2, Yong-Moon Lee1, Jae Hyuk Yi4, Young Jun Ham2,3, Hyung Chul Rah3, Seung Ryul Kim3, Hyung Geun 1,3 Song * 1 Department of Pathology, College of Medicine, Chungbuk National 2 3 University, Cheongju, Research Institute, DiNonA Inc, Iksan, Graduate School of Health Science Convergence, College of Medicine, Chungbuk National University, Cheongju, 4MedClaris Inc, Seoul, Korea Tel: 043-261-2853, E-mail: [email protected] JL1, a specific epitope on CD43, is a potential biomarker for the diagnosis of acute leukemia. Although qualitative assays for detecting leukemia-specific CD43 exist, there is a need to develop quantitative assays for the same. Here, we developed two novel monoclonal antibodies (mAbs), 2C8 and 8E10, recognizing different epitopes on CD43. These clones are capable of pairing with YG5, another mAb against JL1 epitope, because they were selectively obtained using sandwich ELISA. Antigens recognized by 2C8 and 8E10 were confirmed as CD43 by western blotting using the CD43-hFC recombinant protein. When expression on various leukemic cell lines was investigated, 2C8 and 8E10 displayed a disparity in the distribution of the epitope. Enzyme assays revealed that these mAbs recognized a sialic acid-dependent epitope on CD43. Using normal thymus and lymph node paraffin-embedded tissues, we confirmed a difference in the epitopes recognized by the two mAbs that was predicted based on the maturity of the cells in the tissue. In summary, we developed and characterized two mAbs, 2C8 and 8E10, which can be used with YG5 in a sandwich ELISA for detecting leukemia-specific CD43. Keywords: Leukemia, CD43, Epitope, Enzyme-Linked Immunosorbent Assay, Diagnosis Abbreviations: ELISA, enzyme-linked immunosorbent assay; mAb, monoclonal antibody; ALL, acute lymphoblastic leukemia; AML, acute myeloid lymphoma; CHO, chinease hamster ovary Endoplasmic Reticulum Stress Increased Immunosuppression of MDSCs in the Tumor Microenvironment Bora Lee, Bo-eun Kwon, Hyun-jeong Ko* Laboratory of Microbiology and Immunology, College of Pharmacy, Kangwon National University, Chun-cheon, Korea Tel: 033-250-6923, E-mail: [email protected] Although the role of endoplasmic reticulum (ER) stress in cancer has been studied in depth, ER stress is known to increase apoptosis of tumor cells and thus reduce tumor growth. In our study, persistent ER stress induced by multiple administrations of low-dose thapsigargin (Tg) accelerated tumor growth in mice. Tg-mediated ER stress increased the generation of Ly6G+CD11b+ myeloid cells, but did not alter anti-tumor effector T cells. 4-Phenylbutyric acid (PBA), a chemical chaperon which was widely used as ER stress reducer attenuated Tg-induced myeloid-derived suppressor cell (MDSC) expansion and tumor growth. Tg-mediated ER stress enhanced the immunosuppressive capacity of tumor-infiltrating MDSCs by increasing expression of ARG1, iNOS, and NOX2, although splenic MDSCs were not affected. Consistent with these results, 4-PBA restored the anti-tumor immune response by regulating inflammatory cytokines such as TNF-αand CXCL1/KC, and activated tumor-infiltrating CD8+ T cells that were inhibited by Tg-mediated ER stress. These results suggest that significant ER stress in a tumor-bearing host might induce tumor growth mediated by enhancement of MDSC-mediated suppression. Therefore, ER stress reducers such as 4-PBA could restore anti-tumor immunity by inhibiting suppressive MDSCs that are exacerbated by ER stress. Keywords: ER stress, MDSCs, Thapsigargin Characterization of Two Novel mAbs Recognizing Different Epitopes on CD43 P-176 Tumor-derived Osteopontin Suppresses Anti-tumor Immunity by Promoting Extramedullary Myelopoiesis Insu Jeon1, Eun-Kyung Kim2, Hyungseok Seo1, 2 1 2,4 Young-Jun Park , Boyeong Song , Kyoo-A Lee , 1,3 2 2 Yongwoo Jang , Yeonseok Chung , Chang-Yuil Kang * 1 Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence, 2Laboratory of Immunology, Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University Science and Technology, Seoul National University, Seoul, Korea, 3Current address: McLean Hospital, Harvard Medical School, 4 Belmont, MA 02478, Current address: Department of Immunology, College of Medicine, Mayo Clinic, Rochester, MN, USA Tel: 02-880-7860, E-mail: [email protected] Extramedullary myelopoiesis occurs commonly in tumor-bearing animals and is known to lead to accumulation of peripheral myeloid derived suppressor cells (MDSC), which play an important role in immune escape. However, the cellular and molecular mechanisms by which tumors induce extramedullary myelopoiesis are poorly understood. In this study, we found that osteopontin expressed by tumor cells enhances extramedullary myelopoiesis in a CD44-dependent manner through the Erk1/2-MAPK pathway. Osteopontin-mediated extramedullary myelopoiesis was directly associated with increased MDSCs in tumor-bearing hosts. More importantly, osteopontin silencing in tumor cells delayed both tumor growth and extramedullary myelopoiesis, while the same treatment did not affect tumor growth in vitro. Lastly, treatment with an antibody against osteopontin inhibited tumor growth and synergized with cell-based immunotherapeutic vaccines in mediating anti-tumor immunity. Our findings unveil a novel immunosuppressive role for tumor-derived osteopontin and offer a rationale for its therapeutic targeting in cancer treatment. Keywords: Osteopontin, Myeloid derived suppressor cells (MDSC), Tumor, Extramedullary myelopoiesis 102 The 2014 Fall Conference of the Korean Association of Immunologists Tumor and Transplantation Immunology P-177 α-Enolase Stimulates the Cancer Cell Proliferation via TGF-β Production P-178 Poster Presentation NSrp70 Counteracts with SR proteins SRSF1 and SRSF2 and Regulates CD44v5 Alternative Splicing Mirim Jang, Jiyea Choi, Hyemin Kim, Yejin Kim, Jiwon Choi, Jane Jeon, Junman Hong, Jae Seung Kang*, Wang Jae Lee* Chang-Hyun Kim, Hye-Ran Kim, Bo-Ra Na, Min-Sung Kwon, Hyun-Su Lee, Eun-Kyung Choi, Chang-Duk jun* Department of Anatomy, Seoul National University College of Medicine, Seoul, Korea Tel: 02-740-8216, E-mail: [email protected] School of Life Sciences, Immune Synapse Research Center and Cell Dynamics Research Center, GIST, Gwangju, Korea Tel: 062-715-2567, E-mail: [email protected] It has been recently reported that α-enolase (ENO1) is involved in multiple functions such as glycolysis, cancer metastasis and tumor growth. ENO1 is ubiquitously is expressed both in the cytosol and on the cell surface in cancer. However, it remains to be elucidated the function of ENO1 expressed on cancer cells. Thus, we investigated the role of ENO1 in the various cancer cells. First, it was examined the expression of ENO1 on the cell surface in gastric carcinoma cell line, SNU16, colon cancer cell line, HCT116 and lymphoma cell line, U937. As a result, HCT116 and SNU16 slightly expressed ENO1 on their surface, and ENO1 was highly expressed on the U937 cell surface. To identify whether ENO1 is related to cancer cell proliferation or viability, we performed CCK-8 assay after ENO1 stimulation. ENO1 simulation by anti-ENO1 antibody induced the proliferation of all cancer cells. Since tumor growth factor (TGF)-β is known to regulate cellular proliferation and differentiation in cancer, the level of TGF-β was measured, and its level were increased by ENO1 stimulation. These results suggest that ENO1 on the cancer cell surface is involved in the up-regulation of TGF-βproduction and cell proliferation. Nuclear speckle as sub-nuclear membraneless domain is characterized by the assembly of snRNP and SR proteins. The nuclear speckle proteins have been well known to modulate transcription and pre-mRNA processing; hence, other researchers suggested that nuclear speckle serve as storage sites of the pre-mRNA splicing machinery. We have previously reported that Nuclear Speckle-related protein 70 (NSrp70) as novel SR-related protein colocalized with SRSF1 (ASF/SF2) and SRSF2 (SC35) as classical SR proteins and also had function of alternative splice site selection targeting various pre-mRNAs. In the present study, we demonstrate that NSrp70 can regulate CD44 variable 5 exon (v5) alternatives slicing but counteracts with two above SR proteins. Interestingly, deletion of the first predicted RS region (RS1) markedly abrogated the binding to the SR proteins as well as the target mRNA. Accordingly, RS1 deletion mutant also failed to induce alternative splicing. We found that NSrp70 exists as a dimer or multimers that are mediated by coiled-coil (CC) regions locating at the N-terminal region. Mutant analysis revealed that CC domain does not provide the binding sites for SR proteins and target mRNA but is important for the alternative splicing. In conclusion, these results demonstrate that NSrp70 regulates the alternative splicing of CD44v5 via two major domains; one for the dimeric or multimeric interactions and another for binding to the target mRNA. Further study is now processing to understand how NSrp70 counteracts with SRSF1 and SRSF2. Keywords: ENO1, alpha Enolase, TGF-beta Keywords: NSrp70, SR protein, alternative splicing, CD44v5 P-179 Chemoprevention of Azoxymethane/Dextran Sodium Sulfate-Induced Mouse Colon Carcinogenesis by Processed Aloe Vera Gel Heesuk Kim, Jaehee Lee, Chan-Su Park, Sun-A Im, Chong-Kil Lee* College of Pharmacy, Chungbuk National University, Cheongju, Korea Tel: 043-261-2826, E-mail: [email protected] The effects of processed Aloe vera gel (PAG) on mouse colon carcinogenesis were examined in vitro and in vivo. The chemopreventive effect of PAG was examined on azoxymethane(AOM)-initiated and dextran sodium sulfate(DSS)-promoted colon carcinogenesis model in Balb/c mice. Mice were injected i.p. with AOM (10 mg/kg). After 1 week, 2.5% DSS was administered in the drinking water for 7 days followed by 14 days of tap water for recovery, and this cycle was repeated 3 times to establish carcinogenesis. PAG was orally administered at 200 or 400 mg/kg for 13 weeks. The severity of the inflammatory lesions was evaluated by a colon tissue histological assessment. Oral administration of PAG for 13 weeks significantly reduced the multiplicity of colonic neoplasms. Adenoma and adenocarcinoma induced by AOM/DSS treatment was also significantly decreased by the administration of PAG. In conformity with these results, PAG administration reduced the myeloid-derived suppressor cell (MDSC) number in the spleens of colon cancer-bearing mice. PAG was also shown to decrease the percentage of MDSCs when added to cultures of spleen cells containing MDSCs. These findings show that PAG suppresses colitis-related colon carcinogenesis. P-180 Tumor Clone Engineered to Express IL-17A as Membrane-Bound Form Behaves Differently from That Expressed as Secretory Form in vivo Do Thi Van Anh, Young Sang Kim* Department of Biochemistry, College of Natural Sciences, Chungnam National University, Korea Tel: 042-821-7523, E-mail: [email protected] Interleukin-17 is a CD4+ T cell derived proinflammatory cytokine. We investigated the effects of locally produced IL-17A by tumors, either as secretory form or membrane-bound form, to evaluate its biological function. The expression vectors of secretory or membrane-bound forms of IL-17A were transfected into MethA fibrosarcoma, and selected with G418-containing medium. The expression of IL-17A was analyzed by ELISA or RT-PCR using IL-17A specific monoclonal antibody or specific primers. Transduction of IL-17A gene into tumors did not affect in vitro proliferation and expression of MHC class I, L-d. When the tumor clones were in vivo transferred, the IL-17A transfectant of secretory form grew more rapidly when compared with controls. However, the transfectant of membrane-bound form grew slowly in vivo. These results strongly suggest that the membrane-bound form of IL-17A on tumor cells may affect tumor microenvironment with different mode compared with that by the secretory form of IL-17A. Keywords: IL17A, Meth A, Membrane-bound form, Secretory form, Tumor Keywords: Processed Aloe vera gel, Azoxymethane/dextran sodium sulfate-induced colon carcinogenesis, Myeloid-derived suppressor cells The 2014 Fall Conference of the Korean Association of Immunologists 103 Poster Presentation P-181 Tumor and Transplantation Immunology Determination of Functional p53 Domain responsible for the Mitochondria-mediated Apoptosis by Simvastatin P-182 Sang Min Park, Young Sang Kim* Department of Biochemistry, College of Natural Sciences, Chungnam National University, Korea Tel: 042-821-7523, E-mail: [email protected] Simvastatin is an effective cholesterol-lowering drug that has pleiotropic effects including cytotoxicity to cancer cells. We previously reported that simvastatin triggered the mitochondrial apoptosis pathway in MethA fibrosarcoma cells, which was accompanied by translocation of stabilized p53 to the mitochondria. Moreover, eIF2α phosphorylation played a role in cell survival by counteracting the p53-mediated mitochondrial apoptosis in response to statins. In this study, we attempted to identify the functional domain of human p53 protein related to the stabilization and translocation to mitochondria in response to simvastatin. For that purpose we generated five functional domain-deletion mutants of p53 cDNA and subcloned into pCMV-tag2B and pEGFP-C2 vectors. When the pEGFP-C2 vectors containing mutant form of p53 were transfected into 293T cells, the mutants missing NLS (nuclear localization sequences) all were expressed in cytoplasm (mutant 1, 2 and 3). The mutants with NLS were expressed in nuclear only (mutant 4 and 5). This result demonstrates that the NLS sequence is functional to lead the protein to migrate into nucleus. In the HCT116 human colorectal cancer p53 knock out cells, most of the transfectants with pEGFP-C2/mutant p53 except mutant 2 showed reduced apoptosis compared to the wild type p53 transfectant in response to simvastatin. This result indicate that expression of DNA binding domain of p53 may be related to p53-mediated mitochondrial apoptosis pathway in the simvastain-induced apoptosis. Keywords: Statin, p53, Apoptosis A Multimeric Carcinoembryonic Antigen Signal Inhibits the Activation of Human T Cells by a SHP-Independent Mechanism: a Potential Mechanism for Tumor-Mediated Suppression of T Cell Immunity 1 2 2 Eun-Ah Bae , Kyoo-A Lee , You Chan Song , Eun-Kyung 2 2 3 Kim , Yoon-Sook Lee , Tai-Gyu Kim , Chang-Yuil Kang1,2* 1 WCU Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, Seoul National 2 University, Laboratory of Immunology, Research Institute of Pharmaceutical Sciences, College of Pharmacy, 3Department of Microbiology and Immunology, College of Medicine, The Catholic University of Korea, Seoul, Korea Tel: 02-880-7860, E-mail: [email protected] Carcinoembryonic antigen (CEA) is a well-known tumor antigen that is found in the serum of patients with various cancers and is correlated with an increased risk of cancer recurrence and metastasis. To understand the tumor environment and to develop anti-tumor therapies, CEA has been studied as an antigen to activate/tolerate specific T cells. In this study, we show that CEA can function as a co-inhibitory molecule and can inhibit the activation of human PBMC-derived T cells. Addition of CEA-overexpressing tumor cells or immobilized CEA dampened both cell proliferation and the expression of IL-2 and CD69 expression in T cells after TCR stimulation. The phosphorylation of ERK and translocation of NFAT were hampered in these cells, while the phosphorylation of proximal TCR signaling molecules such as ZAP70 and PLCγwas not affected by immobilized CEA. To determine the relevance of CEACAM1 and SHP molecules to CEA-mediated suppression, we tested the effect of the SHP inhibitor, NSC-87877 on CEA-mediated suppression of T cells, but it did not reverse the effect of CEA. Collectively, these results indicate that CEA can function as a modulator of T cell responses suggesting a novel mechanism of tumor evasion. Keywords: CEA, Adhesion molecule, Co-inhibitory signal, Tumor associated antigen, TCR signaling P-183 Reduction in Tumorigenicity of Colorectal Carcinoma Using Plant-Derived GA733-2 Tumor Antigen Ga-Un Lee1, Ha-Rim Choi2, Hyung-Sik Kang1* 1 School of Biological Sciences and Technology, Chonnam National University, 2Department of nursing science, Nambu University, Gwangju, Korea Tel: 062-530-2195, E-mail: [email protected] GA733-2 is colorectal carcinoma (CRC)-associated antigen, which is highly expressed on the cell surface of human colorectal carcinoma. To investigate effect of GA733-2-Fc administration on tumorigenicity of colorectal carcinoma, plant-derived GA733-2-Fc was pre-administered orally to mice at daily intervals. After 2 weeks, MC38 colorectal carcinoma cells were injected and tumorigenicity was evaluated. The final tumor volume and weight were markedly reduced in mice pre-administered with plant-derived GA733-2-Fc. The frequency of apoptotic cells and expression of pro-apoptotic molecules, such as caspase and Bax, were increased in tumor of mice pre-administered with plant-derived GA733-2+ + Fc. The recruitment of activated CD4 , CD8 T cells and NK cells was also enhanced by pre-administration of plant-derived GA733-2-Fc in tumor region. These results implicated that plant-derived GA733-2-Fc prevents the progression of colorectal carcinoma and its application to cancer immunotherapy may be valuable to ameliorate tumor-specific immunity. Keywords: GA733-2, Plant-derived oral vaccine, Colorectal cancer, CD4+ and CD8+ T cell, Natural killer cell P-184 Enhancement of Tumor-Specific T cell Mediated Immunity in Dendritic Cell-Based Vaccines by Mycobacterium tuberculosis Heat-Shock Protein X Seung Jun Lee, Moon Hee Lee, Gun Young Yoon, Da Rae Kang, Tae Heung Kang, Hee Dong Han, Yeong Min Park*, In Duk Jung Department of Immunology, School of Medicine, Konkuk University, Chungju, Korea Tel: 02-2049-6158, E-mail: [email protected] Despite the potential for stimulation of robust antitumor immunity by dendritic cells (DCs), clinical applications of DC-based immunotherapy are limited by the low potency in generating tumor antigen-specific T cell responses. Therefore, optimal conditions for generating potent immunostimulatory DCs that overcome tolerance and suppression are key factors in DC-based tumor immunotherapy. In this study, we demonstrate that the use of the Mycobacterium tuberculosis Heat-shock protein X (HspX) as an immunoadjuvant in DC-based tumor immunotherapy has significant potential in therapeutics. In particular, the treatment aids the induction of tumor-reactive T cell responses, especially tumor-specific cytotoxic T lymphocytes (CTLs). The HspX protein induces DC maturation and pro-inflammatory cytokine production (TNF-α, IL-1β, and IL-6) through TLR4 binding partially mediated by both the MyD88 and TRIF signal pathways. We employed two models of tumor progression and metastasis to evaluate HspX-stimulated DCs in vivo. The administration of HspX-stimulated DCs in+ + creased the activation of naïve T cells, effectively polarizing the CD4 and CD8 T-cells to secrete IFN-γ, as well as enhanced the cytotoxicity of splenocytes against HPV-16 E7 (E7)-expressing TC-1 murine tumor cells in therapeutic experimental animals. Moreover, the metastatic capacity of B16-BL6 melanoma cancer cells toward the lungs was remarkably attenuated in mice that received HspX-stimulated DCs. In conclusion, the high therapeutic response rates with tumor-targeted Th1-type T cell immunity as a result of HspX-stimulated DCs in two models suggest that HspX harnesses the exquisite immunological power and specificity of DCs for the treatment of tumors. This study was supported by a National Research Foundation of Korea (NRF) grant funded by the Korean Government (NRF-2012R1A2A1A03008433). Keywords: Mycobacterium tuberculosis, HspX, Dendritic cell, Cancer vaccination 104 The 2014 Fall Conference of the Korean Association of Immunologists Tumor and Transplantation Immunology P-185 The Regulation of The Pathogenesis of Pancreatic Cancer: Effect of IL-22 and α-Enolase on Human Pancreatic Cancer Cell Line, Panc1 Jiwon Choi, Haemin Kim, Yejin Kim, Mirim Jang, Jiyea Choi, Jane Jeon, Jaeseung Kang*, WangJae Lee* Department of Anatomy, College of Medicine, Seoul National University, Seoul, Korea Tel: 02-740-8294, E-mail: [email protected] IL-22 is a member of IL-10 family, produced by activated DC, T cells, Th17 cells and NK cells. Binding of IL-22 to it’s receptors IL22R1 and IL-10R2 influences variety of immune responses. The receptors are found in non-immune cells such as liver, colon, skin and pancreas. Also the tumours originated from these sites have over expressed IL-22R. Although acinal population of Pancreas has the highest expression of IL22R, Regulation of IL-22 and activation of IL-22 receptor in the pancreatic cancer are poorly understood. In pancreatic cancer cell line, Panc1, introduction of rIL-22 suppressed cell proliferation and induces G2/M phase cell cycle arrest. However IL-22 influenced cytokine TGF-b was increased upon rIL-22 treatment. On the other hand, pro-inflammatory cytokine IL-22 played dual role of anti-tumour effect as well as pro-tumour effect. We have also found the involvement of α-Enolase in the regulation of IL22 and IL22 activation in Panc1 cell line. a-Enolase is known to be highly expressed in cell surfaces of inflammation and cancer and according to clinical research, α-enolase is up-regulated pancreatic cancer patients. Thus it can be suggested that the regulation of IL22 in Panc1 is under the influence of α-Enolase and the mechanisms involved need to be further investigated. P-186 Poster Presentation Involvement of Splenic Lymphoid Tissue Inducer Cells and Regulatory T Cells in Cancer Metastasis Induced by B16-F1 Melanoma Cells Woong-Jae Jung, Ye Eun Lee, Soochan Kim, Hyung Soo Lee, Mi-Yeon Kim* Department of Bioinformatics and Life Science, Soongsil University, Seoul, Korea Tel: 02-820-0458, E-mail: [email protected] In this study, involvement of lymphoid tissue inducer (LTi) cells and regulatory T cells of splenic lymphocytes in cancer metastasis was investigated. After B16-F1 melanoma cells are injected retro-orbitally into a C57BL/6 mouse, eye cancer developes in 2 weeks and metastasizes to lung in some cases. We examined immune responses in spleen during metastasis, and divided into five stages of lung cancer according to the amount of the B16-F1 melanoma cells in lung. The group which developed lung cancer showed increased cell numbers in lymphocytes, B cells, dendritic cells, and CD8+ T cells and decreased numbers in regulatory T cells than that of the control. The number of LTi cells increased in the lung cancer group and their OX40L and CD30L expression was up-regulated, but the number of regulatory T cells decreased. In addition, the percentage of GITR+ regulatory T cells which were undergoing apoptosis increased in the lung cancer group. It suggests that the increased LTi cells and decreased regulatory T cells are related to the cancer metastasis in spleen, and increased apoptosis of GITR+ regulatory T cells implies that the suppressive function of the cells was downregulated during metastasis. Keywords: Tumor, Metastasis, Lymphoid tissue inducer cells, Regulatory T cells, GITR Keywords: IL22, IL22R1, Pancreatic cancer, Panc1, α-Enolase P-187 Role of Scavenger Receptor Type A Agonist Induces Migration of Dendritic Cells to Secondary Lymphoid Organ and Tumor Lesion in Mice Cho-Hee Kim, Chang Gun Kim, Tae-Eon Kim, Jong-Young Kwak* Department of Biochemistry, College of Medicine and Immune-network Pioneer Research Center, Dong-A University, Busan, Korea Tel: 051-240-2856, E-mail: [email protected] Fucoidan, an agonist of scavenger receptor type A (SR-A), has various biological activities, including activation of immune cells and antitumor activity. In this study, a role of SR-A on the migration of dendritic cells (DCs) was investigated. In murine cancer models, intravenous adoptive transfer of bone marrow-derived DCs (BMDCs) but not SR-A-depleted BMDCs from SR-A KO mice in combination with fucoidan achieved tumor growth inhibition. The systemic administration of fucoidan induced maturation of adoptively transferred BMDCs and endogenous DCs in secondary lymphoid organ of mice. Fucoidan induced up-regulation of CCL3, CCL4, CXCL12, TNF-αand CCL21 in secondary lymphoid organ and tumor lesion. In addition, the expression of CXCL12, TNF-αand CCL21 in spleen and Peyer’s patch was higher in fucoidan and DC co-injected mice than in untreated or fucoidan-injected, DC-injected mice. Moreover, CXCL12, TNF-αand CCL21 was expressed in tumor lesion. The expression levels of CXCL12, TNF-αand CCL21 were not increased by fucoidan in SR-A KO mice. These results suggest that administration of SR-A agonist induces migration of DCs to lymphoid organ and tumor lesion through both maturation of DCs and expression of chemokines. Keywords: SR-A, DCs, Fucoidan, Chemokines P-188 IL-7Rα Contributes to Increase of Wound-Healing Migration as well as Invasion in the Prostate Cancer Min A Seol1,2†, Jin-hee Kim1,2†, Keunhee Oh1,2, Ji Hyun 1 1 1,2 3 Sim , Sunkyung Lee , Dong-Sup Lee , Ilkyu Han , 4 1,2 Ja-Lok Ku , Hang-Rae Kim * 1 Department of Anatomy, 2Department of Biomedical Sciences, 3Department of Orthopaedic Surgery, 4Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea † The first two authors equally contributed to this work. Tel: 02-740-8214, E-mail: [email protected] Metastasis to distant organs is an ominous feature of malignant tumors, and the bone is one of the most common sites for metastatic cancer. Especially tumors arising from the prostate possess an increased propensity to spread to the bone. However, the mechanism of metastasis into the bone was not established yet. IL-7Rα plays a critical role in the development of immune cells. Even though there is a circumstantial evidence to involve in the process of bone metastasis, no studies suggest a verification of a valuable part as a key factor of bone metastasis. To investigate the role of IL-7Rαon bone metastases from prostate cancer, we evaluate the expression of IL-7Rα, wound healing migration, invasion, and sub-signals on the prostate cancer cell line such as PC-3, PC3M, PC-M-MM2, and PC-3/nKR after IL-7 treatment. As a result, the expression of IL-7Rα on the bone-metastatic cell line was increased compared to those of others-type metastatic cell-lines. Not only wound-healing migration but also invasion was increased after IL-7 treatment at the bone-metastatic cell line. And, phosphorylation of both AKT and STAT-5 was also increased in time-dependent manner by IL-7 treatment. Moreover, cells with IL-7Rα over-expression or knock-down were established to determine the role of IL-7Rα on the bone metastasis in detail. As a result, cells with IL-7Rαknock-down grew slower than that with control vector. And cells with IL-7Rα over-expression invaded more than that with control vector. In conclusion, this study presents novel evidence demonstrating that IL-7Rα might be associated with bone metastasis of prostate cancer cells upon IL-7 stimulation, leading to migration and invasion. These results form a theoretical basis for the progression of bone metastatic cancer, and future study is needed to examine the effect of the IL-7R α on bone metastasis using animal models. Keywords: Bone metastasis, IL-7Rα, Prostate cancer The 2014 Fall Conference of the Korean Association of Immunologists 105 Poster Presentation P-189 Tumor and Transplantation Immunology KIR Genotype/HLA -C Ligand Combination in Liver Transplantation 1 1 2 Ki Hyun Park , Ji Hyeong Ryu , Heon Woo Park , 3 Eun-Jee Oh * 1 Department of Biomedical Science, Graduate School, Catholic University 2 of Korea, Department of Medical Science, College of Medicine, Catholic University of Korea, 3Department of Laboratory Medicine, College of Medicine, Catholic University of Korea, Seoul, Korea Tel: 02-2258-1641, E-mail: [email protected] Interaction of killer immunoglobulin-like receptors (KIRs) and human leukocyte antigen class-I (HLA-I) regulates NK function. NK regulation influences on NK alloreactivity in transplantation, and HLA mismatch allows the prediction of transplant rejection. But, the clinical impact of KIR/HLA-C combination between recipient and donor on early acute rejection (EAR) is unclear in liver transplantation (LT). KIR (recipients) and HLA-C genotypes (recipients/donors) were measured by Luminex method in 86 living donor liver transplants. HLA-C mismatch (MM), HLA-C type and KIR genotype (inhibitory KIRs, activating KIRs) were analyzed with EAR and MELD score (Model for End-Stage Liver Disease). Of 86 LT patients, 7 patients had EAR. The frequencies of KIR genotypes in 86 recipients were similar with previous studies as KIR2DL1, 2DL2, 2DL3, 2DL5, 2DS1 and 2DS4 were 100%, 16.3%, 100%, 36%, 33.7% and 93%, respectively. HLA-C genotypes of recipients were as follows C1/C1, n=51; C1/C2, n=24; C2/C2, n=1; C1/-, n=10. The frequency of KIR genotypes (KIR2DL1; 100%, 2DL2; 28.6%, 2DL3; 100%, 2DL5; 42.9%, 2DS1; 42.9%, 2DS4; 85.7%) and HLA-C genotypes (C1/C1; n=3, C1/C2; n=2, C2/C2; n=1, C1/-; n=2) in 7 patients of EAR were not different from those of no EAR patients. In terms of HLA– C MM, of 86 patients (MM 0; n=12, MM 1; n=54, MM 2; n=20), 7 patients with EAR showed MM 0 (n=1), MM 1 (n=5) and MM2 (n=1) and there was no significant difference of EAR (P>0.05). The previous report showed that recipient`s KIR (KIR2DL3 and KIR2DS1) enhanced EAR in the presence of donor C2. However, our study was no patients with EAR (donor C2 type). MELD score was also not related with KIR and HLA-C status in LT patients (p>0.05). This study shows a KIR/HLA-C genotype status in Korean LT patients. We could not confirm the impact of KIR/HLA-C genotypes on early outcome of LT. Further studies with long term follow-up will be needed to demonstrate the role of KIR/HLA status in LT. Keywords: NK, Killer immunoglobulin-like receptors, HLA-C, Early acute rejection, Liver transplantation P-191 Anti-HER2/Neu Antibody Therapy Can Reduce the Populations and Immunosuppressive Activity of MDSCs in Mouse HER2/neu+ Breast Tumor Model Jae Hyeog Choi, Kug Hwan Roh, Hyun Keun Song, SaeGwang Park* Laboratory for Medical Oncology, Department of Microbiology and Immunology, College of Medicine, Inje University, Busan, Korea Tel: 051-890-8702, E-mail: [email protected] Myeloid-derived suppressor cells (MDSCs) are a heterogeneous cell populations that play a critical role in tumor associated immune suppression. MDSCs consist of immature cells with myeloid lineage markers CD11b and Gr1 in mice and accumulate in large numbers in tumor bearing mice including HER2/neu+ breast cancers. We have previously reported that the mechanisms of tumor regression by anti-neu antibody require the both innate and adaptive immunity. However, the effect of anti-neu antibody on MDSC remains still not been studied. This study was conducted to test whether anti-neu antibody treatment can affect the populations and immunosuppressive functions of MDSC. Anti-neu antibody reduced the percentage of MDSC in tumor and spleen within 3 days after treatment. In addition, the number of MDSCs in tumor (day2) was decreased earlier than that in spleen (day3). Immunosuppressive activity of MDSC was also reduced by anti-neu antibody treatment. qRT-PCR analysis revealed that chemokine CCL19 and CCL21a which are associated with MDSC recruitment decreased in tumor at day1 and spleen at day2. To increase the effects of anti-neu antibody on MDSC, we combined 5-Fluorouracil (5-FU) known as depleting MDSCs with anti-neu antibody. Combined treatment of anti-neu antibody and 5-FU enhanced tumor regression comparing single therapy such as anti-neu antibody or 5-FU. Reduction of the populations of MDSC was more increased by combined treatment compared to anti-neu antibody single. In addition, combined treatment reduced the populations of PMN-MDSC and TAM that was not reduced in anti-neu antibody single treatment. It is still remain to be clear whether the synergistic effects of combined therapy was caused by increased reduction of MDSC and TAM or by direct effects on tumor cells of 5-FU. However, 5-Fu might be a good candidate to combine with anti-neu antibody for HER2/neu+ breast cancer patients. Keywords: Myeloid-derived suppressor cells, HER2/neu+ breast cancer, Immunosuppressive activity, 5-Fluorouracil 106 P-190 Highly Efficient Adenoviral Vector-Mediated Gene Transfer into Primary Human Natural Killer (NK) Cells for Immunotherapy Seon Ah Lim1, Il-Kyu Choi2, Tae-Jin Kim1, Chae-Ok Yun2, Kyung-Mi Lee1* 1 Department of Biochemistry and Molecular Biology, Korea University College of Medicine, 2Department of Bioengineering, College of Engineering, Hanyang University, Seoul, Korea Tel: 02-920-6253, E-mail: [email protected] Natural kill (NK) cells are effector lymphocytes that play critical roles in the human innate immunity. They provide an important line of defense against many types of bacteria, viruses and tumors. NK cells comprise 10~15% of human peripheral blood lymphocytes and kill tumor cells by releasing small cytoplasmic granules that cause the target cells to die by apoptosis and cause antibody dependent cytotoxicity (ADCC). Since NK cells kill tumor cells without antigen specificity, they can attack a broad range of tumor cells as well as tumor cells that have escaped from CD8T cells. Even if NK cells are potent cells for immunotherapy, current NK cell-based immunotherapies are limited in curing cancer, presumably due to the combination of poor survival, insufficient tumor trafficking, poor anti-tumor function, and severe apoptosis. One way to overcome these hurdles is to introduce proper genes into NK cells leading to the long lasting NK cell survival and function. However, the efficiency of gene transfer into human primary NK cells was poor and induces high level of cell death. Herein, we established a simple and highly efficient transduction method using a modified adenoviral vector, Ad-k35. While various gene transfer systems, such as retrovirus, protein transduction domain, and nanofection, were not successful, the GFP was expressed in up to 80% of primary NK cells using Adenovirus carrying K35 promotor Ad-k35. In contrast, conventional Ad5 or Ad-RGD transduced less than 1% to 8% of primary NK cells. Using Ad-k35, we delivered an anti-apoptosis gene into the expanded NK cells and observed a significant reduction in NK cell apoptosis. These data indicated that Ad-k35 could be used to deliver genes to human NK cells and consequently provide a new experimental approach for cancer immunotherapy. Keywords: NK cells, Adenovirus transduction, Immunotherapy P-192 Platelet-Activating Factor Enhances Tumor Metastasis via the ROS-dependent Protein Kinase CK2-mediated NF-κB Activation Kyoung-Jin Kim1, Kyung-Deuk Cho1, Kyu Yun Jang2, 1 3 3 Han-A Kim , Hae-Kyoung Kim , Hern-Ku Lee , Hwa-Suk 1 1 1 Bang , Soo-Yeon Jeong , Ji-Young Youm , Suhn-Young Im1* 1 Department of Biological Sciences, College of Natural Sciences, Chonnam National University, Gwangju, 2Department of Pathology, Chonbuk National University Medical School, Jeonju, 3Department of Immunology and Institute for Medical Science, Chonbuk National University Medical School, Jeonju, Korea Tel: 062-530-3414, E-mail: [email protected] Platelet-activating factor (PAF) promotes tumor metastasis via activation of the transcription factor NF-κB. We here investigated the role of the protein kinase CK2 in PAF-induced NF-κB activation and tumor metastasis, given that PAF has been reported to increase CK2 activity, and that CK2 plays a key role in NF-κB activation. PAF increased CK2 activity, phosphorylation and protein expression in vivo as well as in vitro. CK2 inhibitors inhibited the PAF-mediated NF-κB activation and expression of NF-κB-dependent proinflammatory cytokines and antiapoptotic factors. Pretreatment with the antioxidant NAC resulted in a significant inhibition in PAF-induced enhancement of CK2 activity, phosphorylation, and protein expression in vivo as well as in vitro. H2O2 and known reactive oxygen species (ROS) inducers, LPS and TNF-α enhanced CK2 activity, phosphorylation, and protein expression, which was again inhibited by antioxidant. PAF-, LPS- and TNF-α-induced increased CK2 activity, phosphorylation, and protein expression, which were inhibited by p38 inhibitor. PAF, LPS or TNF-αincreased pulmonary metastasis of B16F10, which was inhibited by antioxidants, CK2 inhibitor and p38 inhibitor. Our data suggest that 1) ROS activates CK2 via p38, which, in turn, induces NF-κB activation, and 2) PAF, LPS and TNF-αincrease pulmonary tumor metastasis via the induction of ROS/p38/CK2/NF-κB pathway. Keywords: CK2, NF-κB, PAF, ROS, Tumor metastasis The 2014 Fall Conference of the Korean Association of Immunologists Vaccines and Immune Therapeutics P-193 Functional Modulation of T Cell Subsets by Delivering a Transducible Th17-H-TMD Ilkoo Kim, Sang-Kyou Lee* Department of Biotechnology, College of Life science and Biotechnology, Yonsei University, Seoul, Korea Tel: 02-2123-7415, E-mail: [email protected] Th17-H is generally known as a key metabolic regulator in hypoxic condition. Th17-H is a heterodimer which consists of Th17-Hα, an oxygen-sensing molecule, and constitutively expressed Th17-Hβ. Recently, many studies have investigated the link between immune cell development and cellular metabolism. To modulate the function of endogenous Th17-Hα, we suggested that its transcription modulation form could competitively inhibit T cell subsets development. We constructed a transducible competitive inhibitor of Th17-H, tTh17-Hα-TMD, and induced protein expression in E.coli system. To generate tTh17-H-TMD, Th17-HTMD was conjugated with Hph-1-PTD and purified using His-tagged protein purification system. To verify the transduction efficiency of tTh17-H-TMD, we treated Jurkat T cells with the purified protein and detected the protein with immunoblot analysis. Besides, the protein did not affect cell survival by measuring dehydrogenase activity of cells, indicating that tTh17-H-TMD did not cause adverse effect on T cells. In addition, the level of activation markers, CD69 and CD25, were not changed compared to the control group and the proliferation of cell was not affected by tTh17-H-TMD delivery. These data demonstrate tTh17-H-TMD could be successfully transduced into its target cells and did not affect the general function of T cells. Further studies will support the role of Th17-H in CD4+ T cell subsets development and suggest its inhibitory role in T cell differentiation and function. Taken together, Th17-H could be an attractive target for immune disorders and this suggests a novel way for prevention and treatment of autoimmune diseases. Keywords: Hypoxia Inducible Factor-1, T Cell Subsets, Hph-1-PTD, tHIF-1α-TMD, Autoimmune diseases P-195 P-194 1 Department of Bioscience and Biotechnology, Sejong University, Laboratory Science Division, International Vaccine Institute, 3R & D Center, EyeGene, Seoul, Korea Tel: 02-3408-3765, E-mail: [email protected] 2 Influenza virus can cause worldwide pandemic infection, and vaccination is the most effective measure to control influenza infection. However, lack of the cross-reactivity between pandemic and seasonal influenza virus strains causes a shortage of vaccine supply during pandemics. In this study, we investigated the antigen-dose sparing efficacy of a new vaccine adjuvant system CIA06 on pandemic influenza vaccine. Mice were immunized with influenza vaccine antigen alone or in combination with adjuvants, and the immune responses were assessed. Influenza virus-specific serum IgG and HI antibody titers were significantly increased in mice receiving CIA06-adjuvanted vaccine, and this effect was more prominent with low antigen doses. Assays for splenic cytokine secretion indicated significant induction of Th1-type immune responses in these animals. Addition of alum or CIA05 to antigen raised the protective efficacy against influenza virus, but only CIA06 provided the maximum protection at low antigen doses. Lung histology and cytokine expression analysis indicated that no excessive inflammatory responses were induced by the CIA06-adjuvanted vaccine. These results suggest that CIA06 can be used to achieve antigen-sparing effects during pandemic periods. Keywords: Influenza vaccine, Adjuvant system, CIA06 Crucial Roles of Interleukin-7 in the Development of T Follicular Helper Cells and in the Induction of Humoral Immunity YB Seo1, SJ Im1, H Namkoong1, SW Kim1, YW Choi2, MC Kang2, CD Surh2, YC Sung2* 1 Department of Life Sciences, Pohang University of Science and Technology (POSTECH), 2Intergrative Biosciences and Biotechnology, Pohang University of Science and Technology (POSTECH), Pohang, Korea Tel: 02-2258-7527, E-mail: [email protected] T follicular helper (Tfh) cells are specialized providers of cognate B cell help, which is important in promoting the induction of high-affinity antibody production in germinal centers (GCs). Interleukin-6 (IL-6) and IL-21 have been known to play important roles in Tfh cell differentiation. Here, we demonstrate that IL-7 plays a pivotal role in Tfh generation and GC formation in vivo, as treatment with anti-IL-7 neutralizing antibody markedly impaired the development of Tfh cells and IgG responses. Moreover, codelivery of mouse Fc-fused IL-7 (IL-7-mFc) with a vaccine enhanced the generation of GC B cells as well as Tfh cells but not other lineages of T helper cells, including Th1, Th2, and Th17 cells. Interestingly, a 6-fold-lower dose of an influenza virus vaccine codelivered with Fc-fused IL-7 induced higher antigen-specific and cross-reactive IgG titers than the vaccine alone in both mice and monkeys and led to markedly enhanced protection against heterologous influenza virus challenge in mice. Enhanced generation of Tfh cells by IL-7-mFc treatment was not significantly affected by the neutralization of IL-6 and IL-21, indicating an independent role of IL-7 on Tfh differentiation. Thus, IL-7 holds promise as a critical cytokine for selectively inducing Tfh cell generation and enhancing protective IgG responses. Keywords: IL-7, Vaccine adjuvant, Follicular helper T cell Antigen Dose-Sparing Efficacy and Safety of a Pandemic Influenza Vaccine Adjuvanted with a New Vaccine Adjuvant System CIA06 in Mice Ji In Ryu1, Shin Ae Park1, Seo Ri Wui1, Ara Ko1, Jung 2 2 3 Ah Choi , Man Ki Song *, Kwang Sung Kim , Yang Je 3 1 Cho , Na Gyong Lee * Poster Presentation P-196 Intravaginal Immunization of Flagellin-Adjuvanted Peptide Vaccine Induces Tumor Regression in an Orthotopic Genital Cancer Model 1 1 1 Seol Hee Hong , Vivek Verma , Youn Suhk Lee , 1 1 2 Kwangjoon Jeong , Mi Jin Park , Young Chul Sung , Jae-Tae Lee3, Jung-Joon Min1, Joon Haeng Rhee1, Shee 1 Eun Lee * 1 Clinical Vaccine R&D Center, Chonnam National University, Gwangju, 3 Department of Life Sciences, POSTECH, Gyungnuk, Department of Nuclear Medicine, Kyungpook National University School of Medicine, Daegu, Korea Tel: 061-379-8465, E-mail: [email protected] 2 Cervical cancer, a high incidence female cancer, mostly caused by high-risk human papilloma virus (HPV) infection in cervical mucosa. Immunotherapy targeting HPV-derived tumor antigen has been widely studied. We previously demonstrated that a bacterial flagellin, a strong mucosal adjuvant, enhances tumor antigen (TA)-specific CD8+ T cell responses in a subcutaneous (SC) implantation model. Because surrounding tissues of the tumor can shape the tumor microenvironment and influence therapeutic efficacy, immunotherapy in orthotopic genital cancer model is more appropriate for further applications. In this study, we examined whether intravaginal (IVAG) vaccination of flagellin-adjuvanted E6/E7 peptides-based vaccine could induce therapeutic efficacy showing tumor regression in an orthotopic genital cancer model. Co-administration of flagellin with E6/E7 peptide TA-specific IFN-γ production in genital lymph nodes. Vaccination of flagellin- adjuvanted E6/E7 peptides in tumor bearing mice induces tumor regression and long term survival in TLR-5 dependent manner. The IVAG administration route showed superior anti-tumor effect than intranasal (IN) administration. These results suggest that flagellin could be a potent vaginal adjuvant for peptide-based therapeutic anticancer vaccines by TA-specific immune responses through TLR5 stimulation in cancer immunotherapy. Keywords: Orthotopic model, Cervical cancer, Flagellin, Peptide, Vaginal adjuvant The 2014 Fall Conference of the Korean Association of Immunologists 107 Poster Presentation Vaccines and Immune Therapeutics Cellular Immune Response and Protective Immunity to M. tuberculosis Antigen with a de-O-acylated lipooligosaccharide-based Adjuvant Systems in Mice P-197 1 1 1 Ara Ko , Seo Ri Wui , Ji In Ryu, Ji Eun Han , Sung-Jae Shin2, Sang-Nae Cho2, Kwang Sung Kim3, Yang Je Cho3, 1 Na Gyong Lee * 1 Department of Bioscience and Biotechnology, Sejong University, Department of Microbiology and Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, 3R & D Center, EyeGene, Seoul, Korea Tel: 02-3408-3765, E-mail: [email protected] 2 Tuberculosis is a disease caused by infection with Mycobacterium tuberculosis and one third of the world’s population infected. BCG is the only tuberculosis vaccine, but it is not prevent in adults infection and active pulmonary TB. In this study, we investigated the cellular immune response and protective immunity of a new tuberculosis subunit vaccine combined with de-O-acylated lipooligosaccharide (dLOS)-based adjuvant systems. C57BL/6 mice were immunized intramuscularly three times with three tuberculosis recombinant antigens with dLOS-based adjuvant systems, and the immune responses were assessed 4 weeks after last immunization. The tuberculosis subunit vaccine combined with dLOS-based adjuvant systems were induced Th-1 and Th-17 cellular immune responses in splenic and lung cells. Especially, a combination of dLOS and cationic liposomes dimethyldioctadecylammonium (DDA) containing tuberculosis subunit vaccine promote a strong cellular immune responses and increase to Ag-specific CD4+ and CD8+ T cell proliferative potential. Furthermore, it was induced in protective immunity against M. tuberculosis infection. Boosting of BCG-vaccinated mice with dLOS-based adjuvant systems containing tuberculosis subunit vaccines significantly enhances cellular immune responses. These results suggest that dLOS-based adjuvant systems containing tuberculosis subunit vaccine can promote cellular immune response and protective against M. tuberculosis in the mouse model. Keywords: Mycobacterium tuberculosis, Vaccine, Adjuvant, Protection P-199 TREM2 Inhibits Aβ-induced Transcriptional Activation of BACE1 Su-Man Kim1, Ha-Rim Choi2, Hyung-Sik Kang1* 1 School of Biological Sciences and Technology, Chonnam National University, 2Department of nursing science, Nambu University, Gwangju, Korea Tel: 062-530-2195, E-mail: [email protected] TREM2 (Triggering receptor expressed on myeloid cells 2) is a cell surface receptor which play a role in inflammation and development of osteoclast. Previous evidence indicates TREM2 promotes phagocytosis and delays inflammation. It has been also reported that TREM2 mRNA and protein expressions in peripheral blood from Alzheimer’s disease (AD) patients are much higher than in healthy individuals. Microglia in degradation of Ab. the major cause of dementia, is clinically characterized by the extracellular accumulation of Ab. In this study, we demonstrated that TREM2 promoted microglia-mediated amyloid beta (Aβ) clearance through the transcriptional activation of beta-secretase 1 (BACE1) in vitro and in vivo. These results suggest that TREM2 is required for efficient phagocytosis of Aβ and may be a potential target for AD therapies by delaying progression of Aβ. Keywords: TREM2, microglia, phagocytosis, BACE1, Amyloid beta, Alzheimer’s disease P-198 Characterization of the Structure and Immunostimulatory Activity of a Vaccine Adjuvant, de-O-acylated Lipooligosaccharide Ji Eun Han1, Seo Ri Wui1, Kwang sung Kim2, Yang Je Cho2, Wan Je Cho3, Na Gyong Lee1* 1 Department of Bioscience & Biotechnology, College of Bioscience, Sejong University, 2Research & Development Center, EyeGene, 3Yonsei University Gangnam Severance Hospital, Seoul, Korea Tel: 02-3408-3765, E-mail: [email protected] Lipopolysaccharide (LPS) is a major component of the outer membrane of Gram-negative bacteria. LPS elicits strong immunopathological responses during bacterial infection, and the lipid A moiety of LPS is responsible for this immunostimulatory activity. Lipid A exerts its biological activity by sending signals via TLR4 present on immune cells, and TLR4 agonists have been a target for vaccine adjuvant. Previously, we demonstrated an adjuvant activity of deacylated lipooligosaccharide (dLOS) to viral and bacterial antigens. In this study, we characterized the chemical structure of dLOS and evaluated its immunostimulatory activity on mouse and human immune cells in comparison with monophosphoryl lipid A (MPL). dLOS consists of the R3-type core, a glucosamine disaccharide with two phosphate groups, and two N-linked acyl group. dLOS was similar to MPL in induction of cytokine production in mouse peritoneal macrophages, but was a more potent activator in human monocytes and dendritic cells (DCs). Results of an analysis of allogeneic T cell responses revealed that dLOS induces Th1, Th2, and Th17-type immune responses in a dose-dependent manner. The immunostimulatory activities of dLOS were completely abrogated in TLR4(-/-) mice, which confirms its TLR4-dependency. These results suggest that in the presence of the core oligosaccharide, O-linked acyl groups of LPS are dispensable for activating the TLR4 signaling pathway. dLOS did not cause any pathological effects or death at 0.25, 0.5, or 1 mg per kg body weight in mice in the acute toxicity tests. This result suggests that dLOS has a low toxicity. dLOS should be considered for further development as a safe and effective adjuvant for human vaccines. Keywords: de-O-acylated lipooligosaccharide, Vaccine adjuvant, Immunostimulatory activity P-200 Dendritic Cell Based Cancer Immunotherapy Using Antigen and Adjuvant Co-Encapsulated PLGA Nanoparticles Yeong Seon Byeon, Hat Nim Jeon, Eu Gene Baek, Tae Heung Kang, In Duk Jung, Hee Dong Han, Yeong Min Park* Department of Immunology, School of Medicine, Konkuk University, Chungju, Korea Tel: 02-2049-6273, E-mail: [email protected] Dendritic cells (DCs) based cancer immunotherapy requires tumor antigens and adjuvants to be delivered efficiently into DCs. Nanoparticles (NPs) have been developed as carriers for antigen delivery, however, many applications have been limited by the toxicity of matrix, and by the need to use transfection agents to deliver nanoparticles into cells. Here, we developed poly(lactic-co-glycolic acid) (PLGA) NPs encapsulated with ovalbumin as an antigen and poly I:C as an adjuvant, which could effectively deliver antigen into DCs and support DCs maturation. The PLGA-NPs including antigen is efficiently taken up by DCs within one hour and can be detected in vitro by confocal microscopy. Moreover, DCs containing the PLGA-NPs demonstrated higher CD8+ T cell activation and proliferation by cross-presentation compared to control. Mice immunized with DCs containing the PLGA-NPs encapsulated with antigen and adjuvant showed enhanced tumor antigen specific CD8+ T-cell responses in vivo. Administration of PLGA-NPs loaded DCs into EG7 tumor bearing mice was significantly inhibit tumor growth compared to control (p < 0.05). This study demonstrate the potential of PLGA-NPs containing antigen and adjuvant to enhance antigen-specific CD8+ T cell responses, leading that efficacy of PLGA-NPs mediated DC-based tumor immunotherapy is enhanced. This study was supported by the National Research Foundation of Korea (NRF) grant funded the Korea government (NRF-2012R1A2A1A03008433). Keywords: Nanoparticles, Dendritic cell, Cancer immunotherapy 108 The 2014 Fall Conference of the Korean Association of Immunologists Vaccines and Immune Therapeutics P-201 Developing Monoclonal Antibodies Using Recombinant Receptor Binding Heavy Chain Domains of Botulinum Toxin A or B Yu-Ri Kim, Eun Sun Choi, Kiweon Cha, Gi-eun Rhie, Yeon-Hee Kim* Division of High-risk Pathogen Research, Korea National Institute of Health, Korea Centers for Disease Control and Prevention, Chungbuk, Korea Tel: 043-719-8275, E-mail: [email protected] Botulinum neurotoxins (BoNTs), the most potent toxins on earth, are produced from anaerobic and spore-forming Clostridium botulinum, C. baratti, C. butyricum with eight distinct A to H serotypes. Intoxication with BoNTs by ingesting irrelevantly preserved foods can cause the flaccid paralysis of muscles and failure of breadth results in death. Because of the fatal toxicity, BoNTs is known as one of the dangerous agents which can be used as a bioweapon. As it takes several months for the nerve terminal of damaged neuron cells by BoNTs to replenish, early treatment with effective therapeutics can alleviate the aggravation of paralysis and can ultimately shorten the period of hospitalization. However, a licensed therapeutics, equine-derived therapeutic antitoxin is reported to induce side effects and requires high cost. In this study, to develop effective therapeutic monoclonal antibodies, codons of toxin gene were optimized for the efficient expression in E. coli. Thereafter, we made recombinant heavy chain domain of botulinum toxin A or B (rHccA or rHccB) which binds to cellular receptor of neuron cells and used as immunogens. For protection tests, mice were triple immunized and challenged with 1000LD50 of each toxin. All of the immunized mice survived against toxin challenge. To produce mouse monoclonal antibodies which bind to the native heavy chain domain of botulinum toxin A or B, boosting injections were performed with inactivated toxin complexes. Splenocytes of each immunized mouse were fused with myeloma cells and specific antibody-producing cells were screened by ELISA. Efficacy test of toxin neutralization in vivo was performed by mouse bioassay and some clones have shown partial effect of protection against toxin challenge. These results imply that rHccA or rHccB can be used not only as immunogens for developing therapeutic antibodies but for preparing vaccine candidates against naturally occurring or bio-terror botulism cases. Keywords: Botulinum Neurotoxin, Flaccid Paralysis, Therapeutics, Monoclonal Antibody, Mouse Bioassay P-203 Obesity Alters Pulmonary Immune Function and Treating Obesity Improves Mortality against Influenza Infection P-202 Poster Presentation Potency of Pancreatic Adenocarcinoma Up-Regulated Factor (PAUF) as a Novel Adjuvant Protein in Dendritic Cells Vaccine Young Seob Kim, Tae Heung Kang, Yeong-Min Park* Department of Immunology, Lab of Dendritic Cell Differentiation & Regulation, School of Medicine, Konkuk University Tel: 02-2049-6089, E-mail: [email protected] Dendritic cell-based vaccines have been developed for cancer immunotherapy to generate antigen-specific CD8+ T cell immune response. DCs maturation adjuvant without cytotoxicity needs to be improved for Dendritic cell-based vaccine. In this study, we demonstrate that the use of Pancreatic adenocarcinoma up-regulated factor(PAUF) as a novel adjuvant in Dendritic cell-based vaccines has significantly therapeutic efficiency. PAUF induced maturation of dendritic cells (DCs) through TLR4 signal pathway liked LPS as known as TLR4-mediated DCs maturation substance and activated MAPK(p-ERK, p-JNK, p-p38) of TLR4 signal pathway. In addition, this protein increased expression level of pro-inflammatory cytokines (TNF-α, IL-1β, IL6, IL-10, IL-12) and type I IFN-βin vitro. We confirmed that PAUF-mediated matured DCs pulsed with antigen peptide, human papillomavirus type 16 (HPV-16) E7 or OVA, increase the number of antigen-specific CD8+ T cell in C57BL/6 naïve mice. We also showed crucial tumor treatment effect in tumor bearing mice which injected with murine cervical cancer model TC-1, expressing E7, in vivo. In conclusion, PAUF-mediated dendritic cell-based vaccines increase anti-tumor immune response obtained through DCs maturation, activation and antigen peptide presenting. Thus, PAUF, a novel DCs maturation substance, is adequately potential effective immune adjuvant for the cancer immunotherapy. This study was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (NRF-2012R1A2A1A03008433). Keywords: PAUF, Cancer immunotherapy, Dendritic cells P-204 Immunogenicity of Hepatitis A Vaccine in Young Adults Young Jo Song*, Chang Yong Bang, Won Woong Na Joo Young Kim*, Jun Chang Ewha Womans University College of Pharmacy, Seoul, Korea Tel: 02-3277-3016, E-mail: [email protected] Department of Infectious Diseases, Armed Forces Medical Research Institute, Daejeon, Korea Tel: 042-878-4794, E-mail: [email protected] Obesity is a known risk factor for several respiratory diseases. In many countries, obese persons were considered as one of the high risk groups against hospitalizations and mortality during the 2009 influenza pandemic. However, little is known about the mechanism to prove the relationship between obesity and influenza. We assessed general characteristics in obese mice, and whether obesity affects immune functions and mortality during influenza infection. C57BL/6 mice were fed a high fat diet (60% kcal fat) and some obese mice performed voluntary exercise for the treating obesity. The obese mice were immunized with a vaccine based on adenovirus expressing nucleoprotein (rAd/NP) and challenged with influenza virus (PR8). Obese mice showed increased airway hyperresponsiveness (AHR) and collagen levels in the lung parenchyma compared with lean mice. Pulmonary T cell populations were decreased in the obese mice. Especially, naïve T cells were significantly reduced in the lung tissue of the obese mice compared with that of lean mice. In the PR8-challenged experiments, total cytotoxic T lymphocytes (CTLs) were reduced in the lung tissue of the rAd/NP-immunized obese mice compared with the rAd/NP-lean mice. However, NP-specific CTLs were increased in the lung tissue of the obese mice. Both NP-specific antibody titers and mortality rates were similar between rAd/NP-obese and rAd/NPlean mice. Treating obesity recovered the reduced total CTLs of the lung tissue of the rAd/NP-obese mice after PR8 challenge, but NP-CTLs from the treating obesity group still up-regulated. The treating obesity delayed the mortality in the PR8-challenged obese mice. The results demonstrate that obesity changes lung functions-AHR, lung fibrosis and T cell populations in naïve mice. Obesity also changes vaccine effects according to the modifications of the pulmonary CTLs, and treating obesity restore the immune response after influenza challenge. Hepatitis A is an acute infectious disease of the liver caused by the Hepatitis A virus (HAV). Hepatitis A is endemic throughout the world, occurring most commonly, however, in areas of poor hygiene and low socioeconomic conditions. HAV is transmitted via fecal-oral route by the consumption of contaminated water. The age-specific seroprevalence of anti-HAV IgG in Korean children and young adults has decreases as socioeconomic and sanitation conditions improved. Despite introduction of the hepatitis A vaccine in Korea in 1997, this vaccine is not included in the national immunization program. The purpose of this study to evaluate humoral immunity in Korean military personnel after hepatitis A vaccination. A total of 251 trainees voluntarily participated in this study. The mean age of these trainees was 21±1 (range 20-28 years). Approximately 72% of the trainees were 21 years old. Prior to vaccination a blood sample was drawn from all 251 trainees. They were given an adult dose of Epaxal (24IU). Approximately 4 weeks after receiving the vaccine, a second blood sample was collected. The HAV vaccination rate among Korean military recruit trainees was 3.2%. There was no significant difference in the vaccination rate between age groups. Of these 251 trainees, 25 were found to be anti-HAV IgG positive (10%). Four weeks after vaccination, 97.8% of the previously seronegative vaccinees had developed more than 10 mIU/ml of specific antibodies. 7 months after vaccination, 77.3% of vaccinees had more than 10mIU/ml of specific antibodies. In summary, seroprevalence for anti-HAV IgG and HAV vaccination rate of Korean military recruit trainees was low. Seroconversion rate was 97.8% four weeks after vaccination. 7 months after vaccination, 77.3% of vaccinees had more than 10mIU.ml of specific antibodies. It was suggested that the routine HAV vaccination for Korean military personnel might be necessary. Keywords: Adenovirus-based influenza vaccine, Cytotoxic T lymphocytes, Influenza virus, Nucleoprotein, Obesity Keywords: Hepatitis A, Vaccination, Antibody, Immunogenicity The 2014 Fall Conference of the Korean Association of Immunologists 109 Poster Presentation P-205 Vaccines and Immune Therapeutics Human Monoclonal Antibody Binding to Human and Mouse L1CAM Exhibits Antitumor Activity but No Single-dose Toxicity in Mouse Models in Preclinical Studies 1 2 3 Seulki Cho , Haejung Kim , Mooney Lim , Hyun-Kyu 4 1,3 4 Park , Moon Sik Joeng , Woo-Chan Son , Hyo Jeong 1,3 Hong * 1 Institute of Antibody Research, Kangwon National University, 2Department 3 of Biological Science, Kangwon National University, Department of Systems Immunology, Kangwon National University, Chuncheon, 4Department of Pathology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea Tel: 033-250-7363, E-mail: [email protected] L1 cell adhesion molecule (L1CAM) is a 200-220 kDa transmembrane glycoprotein and was first described as a neural cell adhesion molecule. L1CAM is also expressed in malignant tumors and its expression correlates with tumor progression and metastasis. Cholangiocarcinoma is refractory to conventional therapies and overall five-year survival rate is less than ~5 %. Thus, effective therapeutic strategies for cholangiocarcinoma are urgently needed. We previously found that L1CAM plays an important role in tumor progression of cholangiocarcinoma. To evaluate whether anti-L1CAM antibody could serve as an anticancer for the treatment of cholangiocarcinoma, we have developed a human monoclonal antibody Ab417 that specifically binds to human L1CAM (KD, 0.18 nM) and mouse L1CAM (KD, 34.8 pM) with a high-affinity and performed PK, in vivo efficacy and toxicity studies. Half-life of Ab417 was 114.49 ± 10.23 hours when it was administered into rats. Ab417 inhibited tumor growth in a cholangiocarcinoma xenograft nude mice model, and combined treatment with Ab417 and cisplatin exhibited enhanced antitumor activity compared to either agent alone. For a toxicology study, single dose of 10, 30, or 50 mg/kg Ab417 was i.v. administered into ICR mice and after 10 days general symptoms and neuronal toxicity were evaluated. Despite of high affinity to mouse L1CAM, Ab417 exhibited no toxicity in the mice. Taken together, the results suggest that Ab417 has potential as an anticancer agent for the treatment of cholangiocarcinoma. Keywords: L1CAM, Human monoclonal antibody, Cancer, Cholangiocarcinoma Treatment, Antibody engineering P-207 Nanoparticle-forming Polysorbitol Transporter (PST) as an Adjuvant with Subunit Vaccine Enhances Protective Immunity Against Streptococcus pneumoniae Infection 1 1,2 1,3 Yoon Chul Kye , Sung-Moo Park , In Su Cheon , 1 1,2 1 Jannatul Firdous , Byung Shik Shim , Young Jun Ju , Ji Eun Hong1, Man Ki Song2, Seung Hyun Han4, Chong Su 1 1,2 Cho , Cheol-Heui Yun * 1 Department of Agricultural Biotechnology and Research Institute for 2 Agriculture and Life Sciences, Seoul National University, Center for Food and Bioconvergence, Seoul National University, 3Laboratory Sciences Division, International Vaccine Institute, 4Department of Oral Microbiology & Immunology, School of Dentistry, Seoul National University, Seoul, Korea Tel: 02-880-4802, E-mail: [email protected] Vaccines against Streptococcus pneumoniae used today are manly composed of multi-variants of polysaccharides based on their serotypes. However, they have certain limitations including a weak induction of long-term persistency for antigen-specific antibody response and low cross-serotypic protection as there is a lacking of the ini+ tial activity of CD4 helper T cells. On the other hand, for the success of protein subunit vaccine through the mucosal route, development of the appropriate adjuvant is of essence. A number of candidates including cholera toxin and Toll-like receptor ligands such as MPL-A have been evaluated as a potential adjuvant for mucosal vaccination against respiratory pathogens without much success. In this study, we used the polysorbitol transporter (PST), forming nano-sized particles with protein antigens, as a mucosal adjuvant against Streptococcus pneumoniae. Intranasal immunization of the pneumococcal surface protein A (PspA) with PST (PspA-PST) reversed body weight loss and enhanced survival after the lethal dose (50LD50) challenge of S. pneumoniae WU2 strain when compared to the control. The mice immunized with PspA-PST showed higher PspA-specific IgG and IgG1 in the serum, and IgG, IgA in the BAL fluid compared to those of control mice. In addition, these mice showed significantly higher IL-10, but not Th1, Th2 or Th17, responses in the lung and the draining lymph nodes after antigen re-stimulation. Furthermore, until 4 months after the immunization, PspA-specific antibody persisted at a high level in the serum and BAL fluid, and showed protection against the lethal challenge. Taken together, these results suggested that PST can be a potential mucosal adjuvant for S. pneumoniae subunit vaccine. P-206 Modified Outer Membrane Vesicles of Eyedrop and Intranasal Vaccination Protect Against Colonization by Enterohemorrhagic Escherichia coli O157:H7 1 2 3 Ahn Hyun Min , Eun-Do Kim , Sang-Hyun Kim , Kyoung 2 Sub Choi, Kyoung Yul Seo * 1 Department of Ophthalmology, National Health Insurance Corporation Ilsan Hospital, Ilsan, 2Department of Ophthalmology, Eye and Ear Hospital, Severance Hospital, Institute of Vision Research, Yonsei 3 University College of Medicine, Seoul, Viral Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Korea Tel: 02-2228-3581, E-mail: [email protected] Purpose: We sought to examine the immunogenicity and protective efficacy of modified outer membrane vesicles(mOMV) from enterohemorrhagic E. coli O157:H7 via several routes including eyedrop instillation. Methods: BALB/c mice were immunized via several routes including ocular, nasal, sublingual and intramuscular route with mOMVs mixed with cholera toxin(CT). Mice were boosted at 2 weeks. Two weeks after second immunization, mice were challenged with gentamicin resistant-enterohemorrhagic E. coli O157:H7, administered by oral gavage. Before challenge, we quantified serum and mucosal immunoglobulins, and fecal excretions were monitored at 5 days interval by bacterial plating on gentamicin added agar plates after the challenge. Results: Modified OMVs were characterized as having penta-acylated lipid A moiety and did not contain shiga toxin A (STxA) subunit proteins, while retaining the non-toxic STxB subunit. All groups vaccinated with mOMVs with CT elicited greater humoral and mucosal immune responses than PBS-treated groups. Comparisons of E. coli O157:H7 fecal counts among the immunization groups showed significantly reduced counts in eyedrop and nasally administered group. Conclusions: We demonstrated that eyedrop and intranasal immunization with mOMVs with CT stimulate systemic and local antibody responses and reduce EHEC O157:H7 colonization of the small intestine. Keywords: Mucosal vaccine, E. coli, mOMV P-208 Adenoviral mediated Interleukin-10 (Ad:rat IL-10) Gene Transfer in Rat Skin Allograft Lan Sook Chang, Hwayong Ham, youlah Kim, Seok-Chan Eun* Department of Plastic and Reconstructive Surgery, Seoul National University Hospital, Seoul, Korea Tel: 031-787-7223, E-mail: [email protected] Purpose: IL-10 is essential in peripheral tolerance to allergens, autoantigens and transplantation antigens. IL-10 inhibits the expression of costimulators and class II MHC molecules on macrophages and dendritic cells. Because of these actions, IL-10 serves to inhibit T cell activation and terminate cell-mediated TH1 immune reactions. The aims of this study to investigate the immunosuppressive effect of recombinant adenovirus mediated rat IL-10(Ad:rat IL-10) in rat alloskin graft. Methods: The Ad:ratIL-10 was created by inserting the IL-10 cDNA into the pShuttle-CMV vector and was transfected into the 293 cells through liposome mediated vehicles. Donor back skin of Wistar(2X3cm) was grafted to the Sprague-Dawley (SD) recipient. Immuno-suppressive effect of the Ad:rat IL-10 was evaluated by graft survival and compared with control group which was infected by saline. Results: The transfected Ad:ratIL-10 was well documented with viral cytopathogenic effect(CPE) and RT-PCR. Mean grafted survival was 11.5 days in Ad:rat IL-10 group compared to control group of infected with saline(6.6 days, p<0.05). Conclusion: In summary, the Ad:rat IL-10 alone, infected in grafted skin, showed immune-suppressive effect on rat alloskin graft and prolong the graft survival. Keywords: Skin allograft, Recombinant IL-10, Adenoviral vector Keywords: Polysorbitol transporter, Nanoparticle, Mucosal adjuvant, Streptococcus pneumoniae, Protective immunity 110 The 2014 Fall Conference of the Korean Association of Immunologists Vaccines and Immune Therapeutics P-209 Mucosal Immunization with fmOMV and Influenza Vaccine Induces Protection Against Influenza Virus Tae-Young Lee1, Eun-hye Bae1, Doo-Jin Kim1, Chang-Ung Kim1,2, Sang-Hwan Seo1, Sun-Woo Yoon1, Kyu-Tae Chang1, Sang-Hyun Kim1* 1 Viral Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology, 2Department of Biochemistry, Chungnam National University, Daejeon, Korea Tel: 042-879-8276, E-mail: [email protected] Outer membrane vesicles (OMVs), secreted from Gram-negative bacteria, are spherical nano-sized proteolipids enriched with outer membrane proteins and have been utilized in vaccine development. In the present study, we generated a modified OMV (fmOMV) to use it as an adjuvant for the split influenza virus vaccine. We examined the protective efficacy of fmOMV as a mucosal adjuvant for influenza virus vaccine by intranasal immunization. Immunization with mixture of split influenza vaccine antigen and fmOMV induced not only a high antigen-specific IgA response in the lungs and IgG response in serum, including HA- neutralizing antibodies, but also an influenza virus-specific cell-mediated immune response. Group of vaccine antigen with fmOMV protected mice against challenge with lethal dose of the homologous 2009 pandemic H1N1 and heterologous PR8 or H5N2 viruses. These results suggest that the use of fmOMV may be a promising candidate as a safe and potent mucosal adjuvant. Keywords: Mucosal immunzation, OMV, Influenza vaccine P-210 Poster Presentation Bovine Lactoferrin Inhibits the Lethal Septic Shock Induced by Staphylococcal Enterotoxin B (SEB) in Mice Min Jee Kim, Junglim Lee, Seok-Rae Park, Jeecheon Kim1, Yung Choon Yoo* Department of Microbiology, College of Medicine, Konyang University, 1 Agency of Defense Development Tel: 042-600-6491, E-mail: [email protected] Staphylococcal enterotoxin B (SEB) is a bacterial toxin that belongs to the category B of bioterrorism, and causes the lethal shock. Bovine lactoferrin (LF), an iron-binding glycoprotein secreted from neutrophils and epithelial cells in the mammary gland, is known to have diverse biological activities such as antibacterial activity and regulation of immune responses. Here we show the inhibitory effect of LF on SEB-induced lethal shock in mice. In an animal model for SEB-induced lethal shock, Balb/c mice were injected intraperitoneally (i.p.) with SEB (5 ug/mouse), and followed by LPS (lipopolysaccharide, 10 ug/mouse) 3 hr after LPS injection. LF (200 ug/mouse) was treated i.p. 3, 2, and 1 days before SEB injection. Inhibitory effect of LF on SEB-induced septic shock was measured the survival rate of SEB-injected mice 72 hr after SEB treatment. Pre-reatment of LF resulted in a perfect inhibition of SEB-induced septic shock. In addition, treatment of LF significantly inhibited septic shock induced induced by intranasal (i.n) administration of SEB. Treatment of LF significantly reduced the level of cytikines such as TNF-alpha, IL-2, IFN-gamma, and IL-10 in sera of mice treated with SEB. In an ex vivo experiment in which the splenocytes of the mice that had been pre-treated with LF prior to SEB injection were harvested 12 hr after LPS treatment, and re-stimulated with ConA for 48 hr in vitro, it was shown that treatment of LF reduced TNF-a and IFN-g production in ConA-stimulated T cells. This data suggest that LF inhibited SEB-induced septicemia through suppression of T cell function, however, LF did not induce Treg. These results suggest that LF is a promising candidate active for suppression of SEB-induced septic shock. Keywords: Staphylococcal enterotoxin B, Lactoferrin, Septic shock, T cells, Cytokines P-211 Smart Supramolecular Hydrogels encapsulated Bioengineered Stem Cells for Cancer Therapy P-212 Genotyping of MICB Alleles on Microarrays by Improving Allele-Specific Extension Primers Su Jin Kim1, Junseok Yeom2, Byung Woo Hwang2, Hong Namkoong1, Jeonga Yang2, Young Chul Sung1*, Sei 2 Kwang Hahn In-Cheol Baek1, Jung-Pil Jang1, Eun-Jeong Choi2, Tai-Gyu 1,2 Kim * 1 Department of Life Sciences, 2Department of Materials Science and Engineering, Pohang, Korea Tel: 02-2258-7527, E-mail: [email protected] 1 Synthetic hydrogels have been extensively investigated as artificial extracellular matrices (ECMs). Crucial challenges for such hydrogels are sustaining long-term cytocompatible encapsulation and providing appropriate cues for spatio-temporal control of the cells. Here, we report in situ supramolecularly assembled and modularly modified hydrogels for long-term engineered mesenchymal stem cell (eMSC) therapy using cucurbit[6]uril conjugated hyaluronic acid (CB[6]-HA), diaminohexane conjugated HA (DAH-HA), and drug conjugated CB[6] (drug-CB[6]). The eMSCs producing enhanced green fluorescence protein (EGFP) remained alive and emitted the fluorescence within CB[6]/DAH-HA hydrogels in mice for more than 60 days. Furthermore, the long-term expression of mutant interleukin-12 (IL-12M) by eMSCs within the supramolecular hydrogels resulted in effective inhibition of tumor growth with a significantly enhanced survival rate. Taken together, our findings confirm the feasibility of supramolecular HA hydrogels as 3D artificial ECMs for cell therapies and tissue engineering applications. Major histocompatibility complex (MHC) class I chain-related gene B (MICB) plays an important role as a ligand for activating NKG2D that expressed in natural killer (NK) cells, γδT cells, and αβCD8 T cells, and is associated with autoimmune diseases, cancer, and infectious diseases. MICB alleles have been recovered variable single nucleotide polymorphisms (SNPs) to cause human diseases and are discriminated by a variety of methods, including PCR-SBT and PCR-SSP but not microarray-based genotyping. Here, we describe a system for genotyping MICB alleles using allele-specific primer extension (ASPE) on microarrays. Forty high quality, allele-specific extension primers were evaluated using strict and reliable cut-off values for the mean fluorescence intensity (MFI), whereby an MFI >30,000 represented a positive signal and an MFI <10,000 represented a negative signal. Of the eight allele-specific extension primers that were found to be false positives, five were improved by adjusting their length, and three were optimized by refractory modification. MICB genotypes that were identified by ASPE on microarrays showed full concordance with those identified by PCR-SBT. In conclusion, we have developed cost-efficient method for genotyping MICB alleles using ASPE on microarrays, which should be applicable for large-scale SNP typing studies of population and disease associations. Reference 1. Park, K. M.; Yang, J. A. ACS nano 6 (2012) 2960-8. Keywords: Hyaluronic acid, Host-guest interaction, Supramolecular hydrogel, Mesenchymal stem cell therapy Department of Microbiology, 2Hematopoietic Stem Cell Bank, College of Medicine, The Catholic University of Korea Tel: 02-2258-7341, E-mail: [email protected] Keywords: Major histocompatibility complex class I chain-related gene B, Polymerase chain reaction with sequence-based typing, Allele-specific primer extension on microarrays, MICB genotyping The 2014 Fall Conference of the Korean Association of Immunologists 111 Poster Presentation P-213 Technical Innovations and Others Effect of Probiotics Administration on Gut Microbiota Composition 1 3 1 Hye-Ji Kang , HaagLim Cho , Chang-Suk Chae , Ravi 1 1,2 Verma , Sin-Hyeog Im * 1 Academy of Immunology and Microbiology (AIM), Institute for Basic 2 Science (IBS), Division of Integrative Biosciences and Biotechnology (IBB), Pohang University of Science and Technology (POSTECH), Pohang, 3School of Life Sciences, Gwangju Institute of Science and Technology (GIST), Gwangju, Korea Tel: 054-279-2356, E-mail: [email protected] Numerous microbiota live in the gastrointestinal tract of mammalian hosts and it is necessary for the intestinal immune and metabolic homeostasis. The intestinal bacteria are affected by aging process in term of composition, such as core microbiota and diversity level. The gut immune system also undergoes modification in aging and this change may be associated with inflammatory and metabolic disorder. Probiotics are live microorganisms considered to balance microbiota, promote gut health and regulate intestinal homeostasis. In this project, the alteration in the gut microbiota was compared in two groups of mice administered five probiotics mixtures or PBS. The feces samples were collected every week for up 9 to six weeks: PBS or five bacteria (1×10 CFU/mouse), Bifidobacterium bifidum, Lactobacillus acidophilus, Streptococcus thermophilus, Lactobacillus casei and Lactobacillus reuteri were administered daily during three weeks and then washout period was performed for identical duration. Firmicutes and Bacteroides were most predominant phyla of bacteria and Acinobacteria was third major phylum. Bacteroides, Lactobacillus and Bifidobacterium were most abundant genera in Firmicutes, Bacteroides and Acinobacteria respectively. The proportion of Firmicutes was similar with that of week 0 in probiotics group, while it was decreased in PBS group. Bifidobacterium just observed in the early days was represented after the probiotics administration. The continuous probiotics taking seems to lead having the diversity of genera which is decreased by aging. Our study suggests that daily consumption of probiotic can help having the similar microbiota diversity of the young and this may relate to maintain the gut health. This research was supported by Institute for Basic Science (Project cod: IBS-R005-G1-2014-a00). Keywords: Probiotics, Microbiota composition P-215 Flavonoids from Triticum aestivum Inhibit Adipogenesis in 3T3-L1 Cells Sarmila Nepali, Barun Poudel, Hyeon-Hui Ki, Ji-Hyun Lee, Dae-Ki Kim* Department of Immunology and Institute of Medical Sciences, Chonbuk National University Medical School, Jeonju, Korea Tel: 063-270-3080, E-mail: [email protected] This study aims to compare the potential anti-adipogenic effect and underlying mechanism of the flavonoids; luteolin, isoscoparin and isoorientin purified from Triticum aestivum sprout (TA) in 3T3-L1 cells. The cells were treated with different concentrations of flavonoids for 8 days. The lipid accumulation was assessed by Oil-Red-O staining. The expression of transcription factors and genes involved in adipogenesis in the cells was evaluated by real-time reverse transcriptase polymerase chain reaction (qPCR) and western blotting. Results showed that, at 10μM concentration, luteolin, isoscoparin and isoorientin inhibited lipid deposition in the cells by 74, 63 and 65% respectively. The flavonoids also significantly inhibited the transcriptional regulators of adipogenesis such as peroxisome proliferator-activated receptor-gamma (PPARγ), CAAT/enhancer binding protein-alpha (C/EBPα) and sterol regulatory element binding proteins (SREBP)-1c, when compared to the control cells. Likewise, there was significant down-regulation of the adipocyte specific markers associated with lipid metabolism; activating protein (aP)-2, fatty acid synthase (FAS), hormone-sensitive lipase (HSL) and lipoprotein lipase (LPL) in the flavonoids treated cells. Notably, flavonoids treated cells showed enhanced expression of insulin-induced genes (insig-1 and insig-2), which could have blocked activation of adipogenic transcription factor SREBP, eventually leading to adipogenesis inhibition. Taken together, our results show that the flavonoids from TA possess inhibitory effect on adipogenesis through down-regulation of adipogenic transcription factors and genes associated with lipid metabolism, and the upregulation of insig 1 and 2, suggesting that the flavonoids from TA could be potential therapeutic agents for the prevention and treatment of obesity. Keywords: Flavanoids, 3T3-L1 cells, Adipogenesis, PPARγ, C/EBPα 112 P-214 KIAA1114, a Full-length Protein Encoded by the Trophinin Gene, is a Novel Surface Marker for Isolating Tumor-initiating Cells of Multiple Hepatocellular Carcinoma Subtypes 1 2 2 Sae Won Kim , Hyun Gul Yang , Moon Cheol Kang , 2 1 2 Seungwon Lee , Hong Namkoong , Seong Beom An , 2 1,2 Seung-Woo Lee *, Young Chul Sung * 1 Department of Life Sciences and 2Integrative Biosciences & Biotechnology, Pohang University of Science and Technology (POSTECH), Pohang, Korea Tel: 054-279-2294, E-mail: [email protected] Development of tumor-initiating cell (TIC)-directed diagnostics or therapeutics has been challenging owing to the lack of versatile TIC-specific markers that are widely applicable to diverse subsets of heterogeneous human tumors. In this study, we identified a novel hepatocellular carcinoma (HCC)-associated cell surface antigen, KIAA1114, and characterized its role as a unique TIC marker to isolate tumorigenic, stem-cell like populations from both alpha fetoprotein (AFP)-positive and -negative HCC subtypes. KIAA1114 was distinguished from other liver TIC markers by its subtype-independent expression pattern and strict correlation with tumorigenicity in HCC cell lines. Moreover, KIAA1114 expression was strongly detected in primary hepatic tumor, but not in non-tumorous liver high tissues, suggesting its potential clinical diagnostic utility. KIAA1114 + fractions isolated from AFP and AFP- cell lines displayed TIC-like features with superior functional traits than KIAA1114low counterparts, re+ + capitulating known features of EpCAM and CD90 TICs, respectively. Our findings not only address the value of a novel antigen, KIAA1114, as a potential diagnostic factor of human liver cancer, but also as an independent biomarker for identifying TIC populations that could be broadly applied to the heterogeneous HCC subtypes. Keywords: Tumor-initiating cells, Hepatocellular carcinoma, KIAA1114, Cell surface marker P-216 Inflammatory and Cytotoxic effect of Silver Nanoparticles on Human Endothelial Cells Jiyoung Jang, In-hong Choi* Department of Microbiology, Yonsei University College of Medicine, Seoul, Korea Tel: 02-2228-1821, E-mail: [email protected] Due to silver’s antimicrobial properties, silver nanoparticles are used in many biomedical applications. The main objective of this study was to elucidate the cellular effects on endothelial cell include cytotoxicity and cytokine production. The size-dependent cellular effects of silver nanoparticles (5, 100 nm) were evaluated using EA.hy926 cells (human endothelial cell). Silver nanoparticles distribution was characterized by transmission electron microscopy. Cytotoxicity was measured by the cell counting kit-8 assay. The amount of cytokine was quantified using a ELISA Kit. To quantify relative mRNA levels of the genes in EA.hy926 cells, realtime-PCR was performed using a Faststart Universal SYBR Green Master. Whole genome microarray analysis was performed using an Affymetrix GeneChipⓇ Human Gene 2.0 ST Array. 5 nm silver nanoparticles (LD50=1.9 μg/ml) were more cytotoxic on EA.hy926 cells when compared that of 100 nm silver nanoparticles after 24h exposure. After treatment with 5 nm silver nanoparticles IL-11 was released at a concentration greater than 900 pg/ml, and 100 nm silver nanoparticles did not induce IL-11 release. We selected three genes, hemeoxygenase-1 (HO-1), heat shock protein-70 (HSP-70), interleukin-8 (IL-8), and determined their mRNA levels. mRNA levels of genes increased significantly in the 5 nm exposure group. Also, using cDNA microarray, we found that 5nm silver nanoparticles induced the expression of specific genes after exposure. In conclusion, we have found that the silver NPs induce variable extents of cellular toxicity in a dose-dependent and size-dependent manner. And we were able to show that low level exposure to 5 nm silver nanoparticles, but not 100 nm, induces expression of cytokines as well as ROS related genes. This research was supported by NanoㆍMaterial Technology Development Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2009-0082417). Keywords: Silver nanoparticles, Endothelial cell, Microarray, Inflammation, ROS The 2014 Fall Conference of the Korean Association of Immunologists Technical Innovations and Others P-217 Triggering Receptor Expressed on Myeloid Cells 2 (TREM2) Induces High-Fat Diet-Induced Obesity Yun-Hwa Jeong1, Ha-Rim Choi2, Hyung-Sik Kang1* 1 School of Biological Sciences and Technology, Chonnam National 2 University, Department of nursing science, Nambu University, Gwangju, Korea Tel: 062-530-2195, E-mail: [email protected] Triggering receptor expressed on myeloid cells 2 (TREM2) is known to involve in the anti-inflammatory response and osteoclast development. However, the role of TREM2 in adipogenesis or obesity has not yet been defined. Effect of TREM2 on adipogenesis and obesity was investigated in TREM2-transgenic (TG) mice on high-fat diet (HFD). To block TREM2 signaling, a neutralizing fusion protein specific for TREM2 (TREM2-Ig) was used. TG mice were much more obese than wild-type mice after feeding with HFD, independent of the quantity of food intake. These HFD-fed TG mice manifested adipocyte hypertrophy, glucose and insulin resistance and hepatic steatosis. The expression of adipogenic regulator genes, such as peroxisome proliferator-activated receptor γ and CCAAT/enhancer-binding protein α, was markedly increased in HFDfed TG mice. Additionally, HFD-fed TG mice exhibited decreased Wnt10b expression and increased GSK-3β-mediated β-catenin phosphorylation. In contrast, the blockade of TREM2 signaling using TREM2-Ig resulted in the inhibition of adipocyte differentiation in vitro and a reduction in body weight in vivo by downregulating the expression of adipogenic regulators. Our data demonstrate that TREM2 promotes adipogenesis and diet-induced obesity by upregulating adipogenic regulators in conjunction with inhibiting Wnt10b/β-catenin signaling pathway. P-218 Poster Presentation Dissolution Chemistry and Biocompatibility of Single-Crystalline Silicon Nano membranes and Associated Materials for Transient Electronics 1 2 2 Gayoung Park , Suk-Won Hwang , John A. Rogers *, 1 Kyung-Mi Lee * 1 Global Research Laboratory, Department of Biochemistry and Molecular Biology, Korea University College of Medicine, Seoul, Korea, 2Department of Materials Science and Engineering, Frederick Seitz Materials Research Laboratory, University of Illinois at Urbana;Champaign, Urbana, Illinois 61801, United States Tel: 02-920-6253, E-mail: [email protected] Single-crystalline silicon nanomembranes (Si NMs) represent a critically important class of material for high-performance forms of electronics that are capable of complete, controlled dissolution when immersed in water and/or biofluids, sometimes referred to as a type of “transient” electronics. The results reported here include the kinetics of hydrolysis of Si NMs in biofluids and various aqueous solutions through a range of relevant pH values, ionic concentrations and temperatures, and dependence on dopant types and concentrations. In vitro and in vivo investigations of Si NMs and other transient electronic materials demonstrate biocompatibility and bioresorption, thereby suggesting potential for envisioned applications in active, biodegradable electronic implants. Keywords: Silicon nanomembranes, Biocompatible, Biodegradable, Bioresorbable, Transient electronics Keywords: TREM2, Adipogenesis, Adipocyte differentiation, Obesity, Wnt10b/β-catenin P-219 Intravital Imaging of T and B Cell Trafficking Across High Endothelial Venules in Mice Lymph Node P-220 Three-Dimensional Culture of Dendritic Cells and Cancer Cells in Hybrid Nanofibrous Scaffold Kibaek Choe1, Eunjoo Song1, Soyeon Ahn1, Sukhyun 2 2 1,2 Song , Gou Young Koh , Pilhan Kim * Tae-Eon Kim, Chang Gun Kim, Cho-Hee Kim, Jong-Young Kwak* 1 Department of Biochemistry, College of Medicine and Immune-network Pioneer Research Center, Dong-A University, Busan, Korea Tel: 051-240-2856, E-mail: [email protected] Graduate School of Nanoscience and Technology, 2Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Korea Tel: 042-350-1115, E-mail: [email protected] Lymph node (LN) is a major checkpoint for the circulating lymphocytes to recognize foreign antigens. High endothelial venule (HEV) in LN facilitates an effective recruitment of circulating lymphocytes from the blood. HEV has distinctive cuboidal-shaped endothelial cells and prominent perivascular sheaths consisting of fibro-reticular cells (FRCs). Yet, our understanding about cellular dynamics in transendothelial, intra-perivascular channel and trans-FRC migration in HEV has been relatively limited. In this work, we adapted a custom-design intravital microscopy system to visualize T and B cell migration across HEV in real time in vivo. Actin-DsRed transgenic mice were used to visualize HEV-endothelial cells. FRCs were labeled in vivo by injecting anti-ER-TR7 antibody conjugated with far-red fluorophore into the footpad. T or B cells obtained from actin-GFP mice were adoptively transferred. Multiple steps in transendothelial, intra-perivascular channel and trans-FRC migrations of GFP+ T or B cells were successfully imaged. T and B cells squeezed in between endothelial cells and then migrated along perivascular channel, a narrow space between endothelium and FRCs, for searching a proper site to exit by trans-FRC migration. Interestingly, compared with T cells, B cells spent longer time in passing the perivascular channel although their total moving distances in the perivascular channel were similar. By time-lapse imaging during 2 hours, we found there were preferred exit sites (“exit ramp”) from the perivascular channel for both of T and B cell. To observe whether T and B cell exit through the same exit ramp, we simultaneously transferred DsRed+ T and GFP+ B cells into wild type C57BL/6 mice. Indeed, T and B cells followed each other though the same exit ramp from the perivascular channel. In addition to the exit ramp, we also found that there exists an “entrance ramp” to perivascular channel, a preferred site for transendothelial migration for both of T and B cells. Three-dimensional (3D) cultures closely resemble in vivo microenvironment provide new insights into morphological and functional study of immune and cancer cells. In this study, we developed 3D culture system with polycaprolactone nanofibrous scaffolds (NFS) made by electrospinning process. 3D NFS consisted of electrospun nanofibers with diameter ranging from several hundreds to submicron. When dendritic cells (DCs) and cancer cells were seeded on the surface of the NFS, cells infiltrated inside of the NFS. The morphology and spreading of cultured cells in NFS showed are different from those cultured on 2D culture dish. Moreover, activation of DCs and proliferation of cancer cells could be detected in 3D NFS but the pattern of cellular activations in 3D NFS are different from that in 2D culture dish. When DCs were coculture with anticancer drug-treated cancer cells in NFS, DCs sprouted cytoplasm to, synapsed with, and engulfed cancer cells in a 3D manner. We developed NFS that provides 3D structure for functionality of cancer cells and immune cells. Keywords: Cell culture, Cell spreading, Immune response, Polycaprolactone, Scaffold Keywords: Lymph node, High endothelial venule, Lymphocyte, Confocal Microscopy, Intravital Microscopy The 2014 Fall Conference of the Korean Association of Immunologists 113 Poster Presentation P-221 Late Breaking Abstracts ERDR1 Enhances Human NK Cell Cytotoxicity through Degranulation-Dependent Pathway Myungjin Jung, Ha-reum Lee, Daeho Cho* Department of Life Science, Sookmyung Women’s University, Seoul, Korea Tel: 02-710-9416, E-mail: [email protected] Erythroid differentiation regulator 1 (ERDR1) is well known as a stress-related survival factor and exhibits anti-cancer effects against melanoma. However, the function of ERDR1 on immune cells has not been investigated. We studied whether ERDR1 regulates the cytotoxic ability of human natural killer (NK) cells, which are play an important role in innate immune response. In this study, treatment with recombinant ERDR1 resulted in enhanced NK cell cytotoxicity through the secretion of lytic granules. Furthermore, actin modulation was involved in the ERDR1-enhanced NK cell cytotoxicity. ERDR1 stimulated actin accumulation at the immunological synapse, which was induced by the phosphorylation of Vav-1 in NK cells. These findings suggest that ERDR1 may have anti-cancer applications that involve the human immune system. Keywords: Natural killer cell, Erythroid differentiation regulator 1, Cytotoxicity, Degranulation, Actin P-222 Responses of Temperatures for Head, Body, and Rectum of Daily Young Cow According to the Mild Heat Stress Hoyoung Chung*, Yoonjeung Choi, Jiyun Park, Hanul Park Animal Genomics & Bioinformatics Division, National Institute of Animal Science, Suwon, Korea Tel: 031-290-1596, E-mail: [email protected] This study has been aimed to investigate responses of temperatures of head, body, and rectum when exposing heat stress. A total of 10 animals that were aged from 4 to 6 months of age were selected, and heat stress was imposed on animals directly in an environmentally controlled house o that managed 33 C and 90% of humidity during the stress. After exposing heat for 8 hours a day, animals were placed in a normal condition at least 12 hours to recover from the heat stress. The average temperature in the o normal condition was 24 C and 63% of humidity. The analysis confirmed that a significant negative correlation between head and rectum (r=0.647). The correlations between head and body presented r=0.605 while between body and rectum presented the lowest correlations (r=0.391) according to the cumulated heat stress. Thus, heat stress caused unbalanced temperatures among measurements, resulting that heat stress should influence animal performance. The experiment also ascertained that the first heat stress may be cumulated in animal body, and therefore, the next heat stress did not have great impact on animal’s head, body, and rectum temperatures. The final conclusion was that exposing heat stress caused accumulation of several factors that may be maintained with certain levels in the body, and recovering time to 12 hours is not enough for full back of entire immune and organ systems. The immune system for the heat stress should be studied based on biochemical materials in vivo, and genetic networks of immunity for heat stress should be imposed on further analysis. Keywords: Daily cow, Heat stress, Temperature, Immunity P-223 An Interleukin-7 Requirement for iNKT Cell Survival and Homeostasis Principal Investigator Joo-Young Park1, Hilary R. Keller1, Noriko Sato2, 1 Jung-Hyun Park * 1 Experimental Immunology Branch, National Cancer Institute, NIH, Molecular Imaging Program, National Cancer Institute, NIH, USA Tel: 1-301-451-764, E-mail: [email protected] 2 Invariant natural killer T (iNKT) cells are thymus-derived T lineage cells that display phenotypic and functional characteristics of innate NK cells. iNKT cells are critical players in mounting immune responses against foreign antigens, as they can express immediate and high levels of pro-inflammatory cytokines such as IFN-γ and IL-4 upon antigen challenge. Thus, maintaining an intact iNKT cell pool is essential for maintaining effective cellular immunity. Importantly, the cellular factors that control iNKT cell survival and homeostasis are largely unknown. Based on high level expression of the IL-2/15 receptor β-chain on iNKT cells and reduced numbers of iNKT cells in IL-15 deficient mice, iNKT cell survival was thought to be dependent on IL-15 which is a non-redundant cytokine for NK cells. Contrary to this view, here we show that survival and maintenance of iNKT cells were dependent on IL-7 and not on IL-15. Specifically, we found that IL-7-deficient mice showed dramatically reduced iNKT cell numbers both in the thymus and spleen, which suggested that IL-7 is a critical factor for the development and survival of iNKT cells. Moreover, using an in vivo model of peripheral IL-7 deficiency, which we generated by expressing a thymocyte-specific IL-7 transgene in IL-7-deficient mice, we demonstrated that IL-7 is a non-redundant survival factor for iNKT cells in peripheral tissues. In agreement, transgenic overexpression of IL-7 dramatically increased peripheral iNKT cell numbers, whereas overexpression of IL-15 failed to do so. Mechanistically, we found that STAT5 activation downstream of IL-7 receptor signaling was required to induce Bcl-2 expression and to promote survival. Collectively, these results suggest that IL-7, and not IL-15, sets the size of the iNKT cell pool under steady state conditions, and propose that iNKT cells homeostasis is constrained by the same cytokine-dependent mechanism that controls conventional αβ T cell survival and maintenance. Keywords: Homeostasis, Thymus, Cytokines, STAT5, T cells 114 The 2014 Fall Conference of the Korean Association of Immunologists The 2014 Fall Conference of the Korean Association of Immunologists Nov. 6(Thu) ~ 7(Fri), 2014 Seoul, Korea Luncheon Symposium Luncheon Symposium [LS I] Next Generation Technologies for Immuno/Cellular Study and Clinical Application Hak-Jun Ahn Sr. Application Scientist/Business Development, Miltenyi Biotec Korea Miltenyi Biotec is a global provider of products and services that advance biomedical research. From research tools to GMP reagents, Miltenyi Biotec is making the promise of cell therapy a reality. The company’s integrated portfolio of tools supports the translation of basic research into practical applications for human health. Scientists and clinicians around the world use Miltenyi-enabling technologies for effective sample preparation, cell isolation, next generation flow cytometry, New stem cell cell culture media, molecular analysis and preclinical imaging. The area of cell-based therapy in particular requires that all starting materials, including media and cytokines, comply with highly demanding GMP specifications. These qualified products are represented in our rich MACS GMP product portfolio. Ⓡ The MACS brand has set standards in the industry and is trusted across basic, translational and clinical research settings. Right now, even more complex cell selection protocols are available on the next generation CliniMACS ProdigyⓇ device, which provides a fully enclosed and fully automated system, increasing the ease and robustness of Ⓡ complex isolation procedures, making them available to a wide range of users. The MACSQuant Analyzer series provide best-in-class flow cytometry to researchers working in a wide range of fields: from immunology, cell biology, stem cells and pharmacokinetics with fundamental and unique features. For opening new era of fluorescence-based cell sorting, Miltenyi Biotec will introduce new microchip-based cell sorting technology in a fully closed cartridge system. [LS II] Genexine & Technology Won-Seok Lee, Dong-Hoon Choi Genexine The 2014 Fall Conference of the Korean Association of Immunologists 117 Luncheon Symposium [LS III] 재택크의 기본은 세택크와 절세의 방법 소개 사승환 지점장 신한금융 [LS IV] 재택크의 기본은 세택크와 절세의 방법 소개 권정근 본부장 신한금융 118 The 2014 Fall Conference of the Korean Association of Immunologists The 2014 Fall Conference of the Korean Association of Immunologists Nov. 6(Thu) ~ 7(Fri), 2014 Seoul, Korea Author Index The 2014 Fall Conference of the Korean Association of Immunologists Author Index Author Page Session A Author Page Session Byun, Jae Yong 82 P-095 99 P-162 Ahn, Ginnae 60,78 P-005,P-078 Byun, Yong-Soo Ahn, Hak-Jun 117 LS I C Ahn, Huijeong 82, 83 P-096,P-097, P-098,P-099, P-100 Carbone, Francis R. 16 SL IV Cha, Gil Sun 63,79 P-019,P-082 Ahn, Jiyeon 80 P-088 Cha, Hae-Sim 64 P-021 Ahn, Myoung-Hee 87 P-115 Cha, Jeong-Dan 84 P-104 Ahn, Soyeon 56,113 OP 4,P-219 Cha, Kiweon 109 P-201 Ahn, Sun-Young 96 P-151 Chae, Chang Suk 74,112 P-062,P-213 Ahn, Young-Tae 74 P-062 Chang, Jun 54,81,97109 OP 2,P-091, P-153,P-203 Alam, Jehan 61 P-011 Chang, Kyu-Tae 111 P-209 Alfieri, Roberta 19 BS I-1 Chang, Lan Sook 110 P-208 An, Seong Beom 112 P-214 Chang, Sun-Young 100 P-167 Anh, Do Thi Van 103 P-180 Chen, Lieping 54,81 OP 2,P-091 Aoki, Kazuhisa 24 BS II-2 Cheon, In Su 87,110 P-116,P-207 Cheon, Jae Hee 45 BS VII-3 Cho, Bon-A 69 P-044 B Bae, Eun-Ah 104 P-182 Bae, Eun-hye 93,111 P-209,P-139 Cho, Chong Su 110 P-207 Bae, Hyunsu 68,74,77 P-039,P-063, P-073 Cho, Chul-Soo 67,73 P-035,P-058 Bae, Jun-Hyeong 86 P-109 Cho, Chung Hwan 59 P-001 Baek, Eu Gene 108 P-200 Cho, Daeho 75,114 P-065,P-221 Baek, Hyunjung 74 P-063 Cho, HaagLim 112 P-213 Baek, In-Cheol 92,111 P-136,P-212 Cho, Hong R. 88 P-120 Baek, SeungYe 78 P-079 Cho, Hye-Jin 76 P-069 Ban, Chae Yeon 62 P-013 Cho, Hyun-Il 59 P-002 Bang, Chang Yong 109 P-204 Cho, Hyun-Soo 72 P-056 Bang, Hwa-Suk 106 P-192 Cho, Jinhee 60,76,78 P-005,P-006, P-071,P-078 Bing, So Jin 60,76,78 P-005,P-006, P-071,P-078 Cho, Jung-Ah 82 P-094 Boraschi, Diana 19 BS I-1 Cho, Kae Won 59 P-004 Bucala, Richard 43 BS VII-1 Cho, Kyung-Deuk 106 P-192 Byeon, Yeong Seon 108 P-200 Cho, Kyung-Min 68 P-037 The 2014 Fall Conference of the Korean Association of Immunologists 121 The 2014 Fall Conference of the Korean Association of Immunologists Author Page Session Cho, Mi-La 78 P-079 Cho, Sang-Nae 108 P-197 Cho, Seok-Goo 59 P-002 Cho, Seulki 110 Cho, Sunjung Author Index Author Page Session Choi, Jiwon 72,91,103,105 P-053,P-130, P-177,P-185 Choi, Jiyea 72,91,103,105 P-053,P-130, P-177,P-185 P-205 Choi, Jung Ah 107 P-195 72 P-054 Choi, Junhee 66 P-032 Cho, Wan Je 108 P-198 Choi, Kyoung Sub 100,110 P-168,P-206 Cho, Whajung 84 P-101 Choi, Kyung-Ho 69 P-044 Cho, Won-Kyoung 92 P-136 Choi, Kyung-Min 84 P-104 Cho, Woon-Dong 102 P-174 Choi, Min-Young 101 P-172 Cho, Yang Je 107,108 P-195,P-197, P-198 Choi, Mi-Rae 84 P-104 Choi, Sang Yoon 89,91 P-122,P-131 Cho, Yong Mee 102 P-173 Choi, Sang-Yoon 89 P-123 Choe, Jeong Min 66 P-032 Choi, Soo Youn 73 P-058 Choe, Jongseon 71 P-051 Choi, Su Jin 68 P-040 Choe, Kibaek 56,113 OP 4,P-219 Choi, Susanna 73 P-058 Choi, Dong-Hoon 117 LS II Choi, Yeon ho 72 P-054 Choi, Eun Sun 109 P-201 Choi, Yoon Seok 50,65 BS VIII-4,P-028 Choi, Eun-Jeong 92,111 P-136,P-212 Choi, Yoonjeung 114 P-222 Choi, Eun-Kyung 103 P-178 Choi, Young Joon 62 P-013 Choi, Ha-Rim 98,104,108,113 P-157,P-183, P-199,P-217 Choi, Young Ki 53,66 OP 1,P-029 Choi, Hyun jin 94,96 P-141,P-149 Choi, Youngnim 61 P-011 Choi, Il-Kyu 106 P-190 Choi, Yun Ho 62 P-014,P-015 Choi, Inhak 54,80,81 OP 2,P-087, P-091 Choi, Yun Sik 61 P-011 Choi, YW 107 P-194 Choi, In-hong 60,84,92,112 P-007,P-103, P-133,P-216 Chung, Doo Hyun 54,81 OP 2,P-091 Choi, Jae Hyeog 106 P-191 Chung, Hoyoung 114 P-222 Choi, Jae-Hoon 66,85 P-032,P-108 Chung, Hwan-Suck 68 P-039 Choi, Jae-Hyeog 61 P-010 Chung, Hyo Jin 97 P-154 Choi, Je-Min 69,88 P-041,P-119 Chung, Yeon-Ho 60,89 P-008,P-124 Choi, Jeomil 63,79 P-019,P-082 Chung, Yeonseok 102 P-176 Choi, Ji-Hyun 61 P-012 Chung, Yong-Hoon 87 P-115 Choi, Jin-A 87,88 P-113,P-118 Cicatiello, Valeria 67 P-035 122 The 2014 Fall Conference of the Korean Association of Immunologists The 2014 Fall Conference of the Korean Association of Immunologists Author Index Author Cresswell, Peter Page Session 4 PL I D DelProposto, Jennifer L. 59 P-004 Do, Kee Hun 70 P-047 Dong, Chen 42 BS VI-4 Dowling, Mark R. 14 SL III Duffy, Ken R. 14 SL III E Eom, JinBeom 75,88,89 P-066,P-068, P-117,P-121 Eun, Seok-Chan 110 P-208 F Author Page Session Han, Duck-Jong 102 P-173 Han, Eun-Jin 73 P-058,P-059 Han, Hee Dong 104,108 P-184,P-200 Han, Hye-Ju 68 P-038 Han, Ik-Hwan 87 P-115 Han, Ilkyu 105 P-188 Han, Ji Eun 108 P-197,P-198 Han, Ji-Won 77,79 P-075,P-081 Han, Sang-Bae 72,75,88,89 P-055,P-066, P-067,P-068, P-117,P-121 Han, Sang-Seop 87 P-113 Han, Seung Hyun 110 P-207 Falco, Sandro De 67 P-035 Han, Seung-Hyun 87,98 P-116,P-158 Filardy, Alessandra 10 SL I Han, Soo Jung 99,100 P-163,P-168 Firdous, Jannatul 110 P-207 Han, Su-Beam 84 P-104 Frieman, Matthew B. 55,90 OP 3,P-127 He, Jianping 10 SL I Fu, Yang-Xin 65 P-025 Heinzel, Susanne 14 SL III Heo, Kang-Hyuck 77 P-075 G Gang, Sung jun 79 P-084 Ho, I-Cheng 23 BS II-1 Ge, Moyar Q. 55,90 OP 3,P-127 Hodgkin, Philip D. 14 SL III Gebhardt, Thomas 16 SL IV Hong, Bong-Ki 73 P-058 Geletka, Lynn 59 P-004 Hong, Changwan 65 P-026,P-027 Ginhoux, Florent 28 BS III-2 Hong, eun-hye 86 P-110 Hong, Hye-Jin 75 P-065 H Ha, Danbee 60 P-005 Hong, Hyo Jeong 110 P-205 Ha, Sang-Jun 54,81,95 OP 2,P-091, P-147 Hong, Jeong won 102 P-174 Hong, Ji Eun 87,110 P-116,P-207 Ha, Sung-Min 61 P-010 Hong, Jin Tae 72,75,88,89 Ha, Un-Hwan 70 P-045 P-055,P-066, P-067,P-068, P-117,P-121 Haczku, Angela 55,90 OP 3,P-127 Hong, Jinpyo 85 P-107 Hahn, Sei Kwang 111 P-211 Hong, Junman 91,103 P-130,P-177 Ham, Hwayong 110 P-208 Hong, Kwon Pyo 102 P-174 Ham, Young Jun 102 P-174 The 2014 Fall Conference of the Korean Association of Immunologists 123 The 2014 Fall Conference of the Korean Association of Immunologists Author Page Session Hong, Seokmann 102 P-173 Hong, Seol Hee 107 Hong, Sung-Wook Author Index Page Session Jang, Cheol-Hun 79 P-081 P-196 Jang, Eun Kyeong 64 P-023,P-024 100 P-165 Jang, Eunkyung 71 P-049 Hong, Wan-Gi 93 P-137 Jang, Hyung Seok 85 P-108 Hong, Yu Jin 68 P-040 Jang, Jiyeon 90 P-126 Huh, Chul-Sung 74 P-062 Jang, Jiyoung 112 P-216 Hwang, Byung Woo 111 P-211 Jang, Jung-Pil 92,111 P-136,P-212 Hwang, Daehee 67,73 P-035,P-059 Jang, Kiseok 64 P-023 Hwang, Eun Sook 26,80,81 BS II-4,P-086 P-092 Jang, Kyu Yun 106 P-192 Jang, Mirim 72,91,103,105 Hwang, Eun-Ha 93 P-138 P-053,P-130 P-177,P-185 Hwang, Seoung-Hye 67 P-035 Jang, Myoung Ho 29,96 BS III-3,P-152 Hwang, Seung-Mi 84 P-104 Jang, Yong Ho 85,92 P-107,P-135 Hwang, Sue-Yun 73 P-058 Jang, Yong-Suk 98 P-159,P-160 Hwang, Suk-Won 113 P-218 Jang, Yongwoo 102 P-176 Hwang, Sung-Min 74 P-062 Jang, Young-Saeng 84,99 P-103,P-164 Hwang, Won 74 P-062,P-064 Jee, Youngheun 60,76,78 P-005,P-006, P-071,P-078 Hwang, Young Il 72 P-053 Jeon, Eun Su 68 P-040 Hwang, Young-il 91 P-130 Jeon, Hat Nim 108 P-200 Hwang, Yuri 72,90 P-054,P-126 Jeon, Hong Bae 68 P-040 Hyun, Seung-Joo 59 P-002 Jeon, Insu 102 P-176 Jeon, Jane 72,103,105 P-053,P-177, P-185 Jeon, Joo-Hui 94 P-142 Jeon, Jun Ho 89,91 P-122,P-131 Jeon, Jun-Ho 89 P-123 Jeon, Sangjun Davie 78 P-077 Jeong, Euikyong 63,79 P-019,P-082 Jeong, Jae Seok 64 P-022 Jeong, Kwangjoon 107 P-196 Jeong, Min Ho 101 P-170,P-171 Jeong, Soo Kyung 101 P-170,P-171 Jeong, Soo-Yeon 106 P-192 I Iguchi, Tomohiro 24 BS II-2 Im, Sin-Hyeog 74,112 P-062,P-064, P-213 Im, SJ 107 P-194 Im, Suhn-Young 106 P-192 Im, Sun-A 63,103 P-018,P-179 Im, Yu-na 64 P-022 Ishii, Ken J. 20 BS I-2 Italiani, Paola 19 BS I-1 J Jain, Deepika 124 55,90 Author OP 3,P-127 The 2014 Fall Conference of the Korean Association of Immunologists The 2014 Fall Conference of the Korean Association of Immunologists Author Index Author Page Session Jeong, Yu-Jin 88 P-118 Jeong, Yun Hee 54,81,95 OP 2,P-091, P-147 Jeong, Yun-Hwa 113 P-217 Ji, Kon-Young 98 P-157 Ji, Young-Eun 79 Jin, Bo-Ra Author Page Session Jung, Seung-Youn 80 P-088 Jung, Uhee 87 P-114 Jung, Woong-Jae 78,105 P-080,P-186 Jung, Yong Woo 63,69,70 P-020,P-043, P-045 P-081 Jung, Yu-Jin 76,93 P-070,P-137 68,99 P-038,P-164 K Jin, Hyo Sun 91 P-129 Kan, Andrey 14 SL III Jin, Jun-O 97 P-156 Kang, Geun-Hyung 74 P-063 Jin, Shan 74 P-061 Kang, Chang-Yuil 102,104 P-176,P-182 Jo, Eun-Kyeong 38,85,90,91 BS V-4,P-105, P-128,P-129 Kang, Da Rae 104 P-184 Kang, Han Goo 60 P-007 Jo, Hwi-Gyeong 95 P-148 Kang, Ho Sung 63,79 P-019,P-082 Jo, Sangmee Ahn 60 P-005 Kang, Hye-Ji 112 P-213 Jo, Sung-Kee 87 P-114 Kang, Hyung-Sik 98,104,108,113 Jo, Wol Soon 101 P-170,P-171 P-157,P-183, P-199,P-217 Joeng, Moon Sik 110 P-205 Kang, Insoo 69 P-044 Joo, Choun-Ki 99,101 P-161,P-162, P-172 Kang, Jae Seung 72,91,103 P-053,P-130, P-177 Jr. Panettieri, Reynold A. 55,90 OP 3,P-127 Kang, Jaeseung 105 P-185 Kang, Jea-Ran 84 P-104 Ju, HeeJung 99 P-161 Kang, Jung-Ok 97 P-153 Ju, Seong-A 86 P-109 Kang, Kyung-Sun 62 P-016 Ju, Young Jun 110 P-207 Kang, Kyung-Won 77 P-076 Jun, Chang 87 P-116 Kang, MC 107 P-194 Jun, Chang-Duk 74,92,103 P-062,P-134, P-178 Kang, Min-Jung 87 P-113 Jun, Jae-Bum 88 P-120 Kang, Mi-Seon 61 P-010 Jun, Ka-jung 81 P-089 Kang, Moon Cheol 112 P-214 Jung, In Duk 104,108 P-184,P-200 Kang, Seong Wook 60 P-008 Jung, Jae Ung 35 BS V-1 Kang, Seong-Ho 68,99 P-038,P-164 Jung, Keunok 80 P-087 Kang, Seong-Mook 80 P-088 Jung, Kyung-Hwa 74 P-063 Kang, Shin Myung 76 P-072 Jung, Min Kyung 76 P-072 Kang, Su jin 94 P-141 Jung, Myungjin 114 P-221 Kang, Sujin 95 P-146 The 2014 Fall Conference of the Korean Association of Immunologists 125 The 2014 Fall Conference of the Korean Association of Immunologists Author Page Session Kang, Tae Heung 104,108,109 P-184,P-200, P-202 Kang, Tae-Bong 77,79 P-075,P-081 Kang, Wonseok 59 P-001 Kang, Yoon Joong 95 P-145 Kang, Yoon-Joong 94,95 P-142,P-148 Kang, Young Mo 44 BS VII-2 Keller, Hilary R. 114 P-223 Kelsall, Brian 10 SL I Kho, In Seong 63,67 P-017,P-036 Ki, Hyeon-Hui 112 P-215 Kim, Areum 60,76,78 P-005,P-006, P-071,P-078 Author Index Author Page Session Kim, Ga-Young 71 P-050 Kim, Gi-Cheon 74 P-062 Kim, Girak 98 P-158 Kim, Green 87 P-113 Kim, Haejung 110 P-205 Kim, Hae-Kyoung 64,106 P-022,P-192 Kim, Haemin 105 P-185 Kim, Han-A 106 P-192 Kim, Hang-Rae 69,105 P-044,P-188 Kim, Heesuk 103 P-179 Kim, Hyekang 66 P-031 Kim, Hyemin 72,91,103 P-053,P-130, P-177 Kim, Hyeong-Su 73 P-060 Kim, Hyeon-Jin 68 P-038 Kim, Hye-Ran 92,103 P-134,P-178 Kim, Hyoung-Pyo 30,62 BS III-4,P-013 Kim, Hyun Gyung 69 P-043 Kim, Hyun J. 88 P-120 Kim, Hyung-Sik 62 P-016 Kim, Hyunseong 68 P-039 Kim, Hyunwoo 91 P-130 Kim, Ilkoo 107 P-193 Kim, Isaac 90 P-128 Kim, Jae Ouk 96 P-152 Kim, Jeecheon 111 P-210 Kim, Jeeyoung 82,83 P-096,P-097, P-098,P-099, P-100 Kim, Ji Sung 72,75,88,89 P-055,P-066, P-067,P-068, P-117,P-121 Kim, Bo-Kyung 100 P-166 Kim, Bo-Yeon 87 P-113 Kim, Byoung Joon 76 P-069 Kim, Byung-Sam 86,88 P-109,P-120 Kim, Chang Gun 105,113 P-187,P-220 Kim, Chang Hwan 47 BS VIII-1 Kim, Chang-Hoon 97 P-154 Kim, Chang-Hyun 103 P-178 Kim, Chang-Ung 93,111 P-139,P-209 Kim, Cho-Hee 105,113 P-187,P-220 Kim, Da Sol 69,70 P-042,P-046 Kim, Dae Seung 76 P-071 Kim, Dae-Ki 112 P-215 Kim, Do-Hyun 69 P-041 Kim, Dong-Jae 87,88,93 P-113,P-118, P-138 Kim, Dongwook 70,101 P-047,P-169 Kim, Doo-Jin 93,111 P-139,P-209 Kim, Eun-Do 100,110 P-168,P-206 Kim, Ji Young 86 P-112 Kim, Eunhee G. 61 P-009 Kim, Ji-Hae 97 P-155 Kim, Eun-Kyung 102,104 P-176,P-182 126 The 2014 Fall Conference of the Korean Association of Immunologists The 2014 Fall Conference of the Korean Association of Immunologists Author Index Author Page Session Kim, Jihye 53,62,66 OP 1,P-013, P-029 Kim, Jin Kyung 91 P-129 Kim, Jin-hee 69,105 P-044,P-188 Kim, Jinhee 60 P-005 Kim, Jini 71 P-051 Kim, Jiyoung 73,94 P-057,P-144 Kim, Ji-Young 95 P-145 Kim, Jong Hoon 62 P-013 Kim, Jong-Min 79 P-084 Kim, Joo Hee 70 P-048 Kim, Joo Young 109 P-203 Kim, Joong Gon 69 P-044 Kim, Joong Sun 101 P-170,P-171 Kim, Jung Hwan 96 P-152 Kim, Jung-Eun 74 P-062 Kim, Jung-Hyun 87 P-115 Kim, Juyang 88 P-120 Kim, Juyoung 72,75,88,89 P-055,P-066, P-067,P-068, P-117,P-121 Author Page Session Kim, Kyoung-Jin 106 P-192 Kim, Kyu-Hui 95 P-148 Kim, Kyungjae 96 P-150 Kim, Mi Eun 67 P-033 Kim, Mi-Hyoung 80,98 P-088,P-157 Kim, Min Jee 111 P-210 Kim, Min Jung 96 P-152 Kim, Min-Gyu 87 P-113 Kim, Minji 63 P-018 Kim, Minsoo 49 BS VIII-3 Kim, Min-Young 102 P-174 Kim, Mi-Yeon 78,105 P-080,P-186 Kim, Moon Gyo 78 P-077 Kim, Myung Hee 90 P-128 Kim, Nam-Hoon 73 P-058 Kim, Nayoung 34,102 BS IV-4,P-173 Kim, Pilhan 56,113 OP 4,P-219 Kim, Pyeung-Hyeun 68,79,99 P-038,P-083, P-164 Kim, Sae Won 112 P-214 Kim, Kang Mi 69,70 P-042,P-046 Kim, Sae-Hae 98 P-159,P-160 Kim, Ki Yeon 78 P-077 Kim, Sang Hoon 82 P-095 Kim, Kiwan 94 P-143 Kim, Sang-Hyun 93,110,111 P-139,P-206, P-209 Kim, Kristine M. 61 P-009 Kim, SeoHyeong 74 P-061 Kim, Kwang Hee 86 P-112 Kim, Seung Jae 60 P-007 Kim, Kwang Soon 66 P-031 Kim, Seung Ryul 102 P-174 Kim, Kwang Sung 107,108 P-195,P-197, P-198 Kim, Shoo-Eon 59 P-002 Kim, Kwanghee 73,96 P-057,P-150 Kim, Song-Ee 62 P-013 Kim, Kwang-Hee 94 P-142 Kim, Soo Yeon 85 P-105 Kim, Kwangsoon 100 P-165 Kim, Soochan 78,105 P-080,P-186 Kim, Kyeongdae 66 P-032 Kim, Soo-Chan 62 P-013 Kim, Kyong Hoon 70 P-045 Kim, Soseul 102 P-174 The 2014 Fall Conference of the Korean Association of Immunologists 127 The 2014 Fall Conference of the Korean Association of Immunologists Author Page Session Kim, Su Gang 78 P-077 Kim, Su Jin 97,111 Kim, Su-Man Author Index Page Session Kim, Yong-Hee 79 P-084 P-154,P-211 Kim, Yoon-Keun 46 BS VII-4 108 P-199 Kim, youlah 110 P-208 Kim, Sung Dae 101 P-170,P-171 Kim, You-Me 48,66 BS VIII-2,P-031 Kim, Sung-Jo 63,79 P-019,P-082 Kim, Young 74 P-062 Kim, Sun-Jin 79 P-083 Kim, Young Hun 72 P-056 Kim, Suyeon 91 P-129 Kim, Young Min 66 P-031 Kim, SW 107 P-194 Kim, Young Sang 103,104 P-180,P-181 Kim, Tack-Joong 64 P-021 Kim, Young Seob 109 P-202 Kim, Tae Hoon 93,97 P-140,P-154 Kim, Young-In 100 P-167 Kim, Tae Jin 63,67 P-017,P-036 Kim, Young-Joo 82 P-094 Kim, Tae Sung 67,68,71,75,86,91 P-034,P-037, P-052,P-065, P-111,P-129, P-132 Kim, Youngsoo 72,75,88,89 P-055,P-066, P-067,P-068, P-117,P-121 Kim, Youn-Hee 84 P-102 Kim, Tae Wan 72 P-053 Kim, Yu-Ri 109 P-201 Kim, Tae-Eon 105,113 P-187,P-220 Kitamura, Kazuya 10 SL I Kim, Tae-Gyun 30,62 BS III-4,P-013 Ko, Ara 107,108 P-195,P-197 Kim, Tae-Hyoun 87,93 P-113,P-138 Ko, Eun-Sil 84 P-104 Kim, Tae-Jin 65,94,106 P-025,P-144, P-190 Ko, Hyun-Jeong 68,79,86,100,102 P-038,P-083, P-110,P-167, P-175 Ko, Youngho 69 P-044 Kobayashi, Takaaki 33 BS IV-3 Koh, Gou Young 56,113 OP 4,P-219 Kole, Abhi 10 SL I Komori, Kuniharu 81 P-089 Kong, Hyunseok 96 P-150 Koo, Bo Kyung 61 P-009 Koziol-White, Cynthia J. 55,90 OP 3,P-127 Kronenberg, Mitchell 41 BS VI-3 Ku, Ja-Lok 105 P-188 Kumar, Vinay 65 P-025 Kwak, Jong-Young 105,113 P-187,P-220 Kim, Tae-Joo 82 P-094 Kim, Tai-Gyu 59,92,104,111 P-002,P-136, P-182,P-212 Kim, Wan-Uk 67,73 P-035,P-058, P-059 Kim, Woan-Sub 68,79,99 P-038,P-083, P-164 Kim, Won-Ju 88 P-119 Kim, Won-keun 55,90 OP 3,P-127 Kim, Wonyoung 88 P-120 Kim, Yejin 72,91,103,105 P-053,P-130, P-185,P-177 Kim, Yeon-Hee 109 P-201 Kim, Yong Guk 72,75,88,89 P-055,P-066 P-067,P-068, P-117,P-121 128 Author The 2014 Fall Conference of the Korean Association of Immunologists The 2014 Fall Conference of the Korean Association of Immunologists Author Index Author Page Session Kwak, Kyung Il 85 P-107 Kwak, Man Sup 72 Kwok, Seung-Ki Page Session Lee, Gap-Ryol 73,82 P-060,P-093 P-056 Lee, Gapryol 94 P-143 78 P-079 Lee, Ga-Un 104 P-183 Kwon, Bo-eun 102 P-175 Lee, GeonHee 71 P-049 Kwon, Bo-Eun 79,86 P-083,P-110 Lee, Geun-Shik 82,83 Kwon, Byungsuk 31,88 BS IV-1,P-120 P-096,P-097, P-098,P-099, P-100 Kwon, H Moo 73 P-058 Lee, Gihyun 74 P-063 Kwon, Hyeok-Il 53,66 OP 1,P-029 Lee, Ha-reum 114 P-221 Kwon, Joonha 94 P-144 Lee, Ha-Yan 98 P-159,P-160 Kwon, Junghak 96 P-150 Lee, Hemin 74 P-061 Kwon, Min-Sung 92,103 P-134,P-178 Lee, Hern-Ku 64,106 P-022,P-192 Kwon, Ohseop 66 P-031 Lee, Heung Kyu 59,93 P-003,P-140 Kwon, Soon-Jae 68 P-040 Lee, Hong Kyung 72,75,88,89 Kye, Yoon Chul 87,110 P-116,P-207 P-055,P-066, P-067,P-068, P-117,P-121 Lee, Hye-Mi 91 P-129 L Author Le, Hongnga T. 88 P-120 Lee, Hyeonhoon 74 P-063 Lee, Arim 86 P-111 Lee, Hyo-Ji 76 P-070 Lee, Bo-Ra 86,100 P-110,P-167 Lee, Hyun Soo 101 P-172 Lee, Bora 102 P-175 Lee, Hyung Soo 78,105 P-080,P-186 Lee, Byoung-Hee 54,81 OP 2,P-091 Lee, Hyun-Joo 59 P-002 Lee, Byunghyuk 65 P-027 Lee, Hyunju 72 P-054 Lee, Chang Geun 101 P-170,P-171 Lee, Hyun-Su 92,103 P-134,P-178 Lee, Changhon 74 P-062,P-064 Lee, Ik-Rae 87 P-113 Lee, Chanju 77 P-073 Lee, In-Young 98 P-157 Lee, Chong-Kil 63,103 P-018,P-179 Lee, Jaehee 103 P-179 Lee, Choong-Eun 71 P-050 Lee, Jaeseon 78 P-079 Lee, Deuk-Ki 84 P-101 Lee, Jae-Tae 107 P-196 Lee, Dong-Gun 78 P-079 Lee, JaeUng 69 P-041 Lee, Dong-Sup 69,105 P-044,P-188 Lee, Jeewon 50,53,65,66 BS VIII-4,OP 1, P-028,P-029 Lee, Eun Jung 96 P-152 Lee, Jennifer 78 P-079 Lee, Eungchang 80 P-087 Lee, Jeong-Min 79,99 P-083,P-164 Lee, Eunhye 94,95 P-141,P-146 Lee, Ji-Hyun 61,112 P-010,P-215 The 2014 Fall Conference of the Korean Association of Immunologists 129 The 2014 Fall Conference of the Korean Association of Immunologists Author Page Session Lee, Jin-Wook 95 P-145 Lee, Joo Hee 63,67 Lee, Joy G. Author Index Author Page Session Lee, Seung Young 96 P-152 P-017,P-036 Lee, Seungwon 66,112 P-031,P-214 90 P-128 Lee, Seung-Woo 66,97,112 P-031,P-155, P-214 Lee, Jun Ho 94 P-144 Lee, Shee Eun 107 P-196 Lee, Jun Sik 67 P-033 Lee, Shin-Ae 72 P-056 Lee, Jun Young 70 P-048 Lee, Soojin 85 P-107 Lee, Jung Eun 86 P-112 Lee, So-Young 74 P-062 Lee, Jung-Ah 88 P-119 Lee, Suk-Kyeong 59 P-002 Lee, Junglim 80,111 P-085,P-210 Lee, Sun-Deok 59 P-002 Lee, Jungsoo 74 P-061 Lee, Sung Joong 85,92 P-107,P-135 Lee, Junho 73 P-057 Lee, Sungwon 96 P-150 Lee, Kihwa 63 P-018 Lee, Sungwook 94,95,96 Lee, Kwang Hoon 74 P-061 P-141,P-146, P-149 Lee, Kwang-Ho 77,79 P-075,P-081 Lee, Sun-Ho 79 P-084 Lee, Kyoo-A 102,104 P-176,P-182 Lee, Sunkyung 105 P-188 Lee, Kyung-Bok 87,88,93 P-113,P-118, P-138 Lee, Sun-Kyung 69 P-044 Lee, Tae-Young 93,111 P-139,P-209 Lee, Kyung-Mi 32,65,73,86,94, 106,113 BS IV-2,P-025, P-057,P-112, P-144,P-190, P-218 Lee, Taeyun A. 94,95 P-141,P-146 Lee, Wang Jae 72,91,103,105 P-053,P-130, P-177,P-185 Lee, Won-Seok 117 LS II Lee, Won-Woo 60,72,89,90 P-008,P-054, P-124,P-126 Lee, Kyungmin 98 P-158 Lee, Mi Sun 69,70 P-042,P-046 Lee, Mi-Ran 85 P-108 Lee, Mi-Yeon 68 P-040 Lee, Wonyong 73,82 P-060,P-093 Lee, Moon Hee 104 P-184 Lee, Ye Eun 78,105 P-080,P-186 Lee, Myung-Shik 36 BS V-2 Lee, Yea-Sol 86 P-109 Lee, Na Gyong 107,108 P-195,P-197, P-198 Lee, Yong-Moon 102 P-174 Lee, Yoo-Dong 64 P-022 Lee, Sang-Ho 93 P-139 Lee, Yoon-Sook 104 P-182 Lee, Sang-Kyou 107 P-193 Lee, Youn Suhk 107 P-196 Lee, Sang-Myeong 61,77 P-012,P-076 Lee, Youngjoo 96 P-150 Lee, Sang-Yeon 96 P-151 Lim, Dae-Seog 80 P-088 Lee, Sasung 67 P-035 Lim, Hui Xuan 75 P-065 Lee, Seung Jun 104 P-184 130 The 2014 Fall Conference of the Korean Association of Immunologists The 2014 Fall Conference of the Korean Association of Immunologists Author Index Author Page Session Page Session Lim, Ji-ye 84 P-104 N Lim, Mooney 110 P-205 Na, Bo-Ra 92,103 P-134,P-178 Lim, Sangho 88 P-119 Na, Kyung-Sun 99 P-161,P-162 Lim, Seon Ah 73,94,106 P-057,P-144, P-190 Na, Won Woong 109 P-204 Na, Woon-Seong 93 P-139 Lim, Sojung 77 P-074 Nair, Varun Sasidharan 81 P-090 Lin, Xiaotian 6 PL II Nam, Jae-Hwan 84,96 P-101,P-151 Lopez, Carolina B. 55,90 OP 3,P-127 Namkoong, H 107 P-194 Lumeng, Carey N. 59 P-004 Namkoong, Hong 111,112 P-211,P-214 Nepali, Sarmila 112 P-215 Ng, Joanne 94 P-144 Nguyen, Loi T. 90 P-128 Nguyen, Quang-Tam 88 P-120 Nguyen, Tam 86 P-109 Nguyen, Thu-Ha T. 86 P-109 Noh, Ji Yoen 74 P-061 Noh, Kyung Tae 85 P-106 Oh, Dong Sun 59 P-003 Oh, Eun-Jee 106 P-189 Oh, Goo Taeg 85 P-108 Oh, Hana 61 P-010 Oh, In Soo 59 P-001 Oh, Keunhee 69,105 P-044,P-188 Oh, Kwon Ik 81 P-090 Oh, Na-Hyun 77 P-075 Oh, Sang-min 67 P-035 Oh, Sang-Muk 87,88,93 P-113,P-118, P-138 M Author Mackay, Laura 16 SL IV Maine, Christian 6 PL II Mamura, Mizuko 25 BS II-3 Marchingo, Julia M. 14 SL III Markham, John. F. 14 SL III Matthews, Krystal 55,90 OP 3,P-127 Matzinger, Polly 82 P-094 Milanesi, Luciano 19 BS I-1 Min, Ahn Hyun 110 P-206 Min, Byoung-Hoon 79 P-084 Min, EunJu-Hong 76 P-069 Min, Hyun Jin 97 P-154 Min, Hyunjung 85,92 P-107,P-135 Min, Ji-Young 22 BS I-4 Min, Jung-Joon 107 P-196 Miyatake, Shoichiro 24 BS II-2 Mok, Jee-Won 99 P-161,P-162 Moon, Hye-Jung 82 P-094 Moon, U-jeong 88 P-120 Moon, Yoo Ri 102 P-174 Oh, Sejin 77 P-074 Moon, Yuseok 70,101 P-047,P-169 Oh, Sera 81 P-092 Morris, David L. 59 P-004 Oh, Se-Woong 85 P-108 Mosca, Ettore 19 BS I-1 Oh, Su Jung 101 P-170,P-171 O The 2014 Fall Conference of the Korean Association of Immunologists 131 The 2014 Fall Conference of the Korean Association of Immunologists Author Page Session Oh, Wonil 68 P-040 Oh, Yeon Ji 81 P-092 Oh, Yeon Kyung 64 P-023,P-024 Ohteki, Toshiaki 27 BS III-1 P Author Index Author Page Session Park, Jong-Hwan 87,88,93 P-113,P-118, P-138 Park, Joon Seok 54,81 OP 2,P-091 Park, Joo-Young 114 P-223 Park, Jun Bae 72 P-056 Park, Jun Beom 91 P-132 Park, Jung-Hyun 114 P-223 Park, Ki Hyun 106 P-189 Park, Kyong Soo 61 P-009 Paik, Da-Young 87 P-113 Park, Ae Kyung 69 P-044 Park, Areum 95,96 P-146,P-149 Park, Bonggoo 86,94 P-112,P-144 Park, Mi Jeong 75 P-067 Park, Boyoun 94,95,96 P-141,P-146, P-149 Park, Mi Jin 107 P-196 Park, Chan Hee 54,81 OP 2,P-091 Park, Mi-Kyung 78 P-079 Park, Chang Ook 74 P-061 Park, Minhwa 100 P-166 Park, Chanho 63,67 P-017,P-036 Park, Moon Suh 82 P-095 Park, Chan-Su 103 P-179 Park, Ok Kyu 89 P-123 Park, Chung-Gyu 79 P-084 Park, Ok-kyu 89,91 P-122,P-131 Park, Do Yang 97 P-154 Park, SaeGwang 61,106 P-010,P-191 Park, Dong Choon 82 P-095 Park, Sang Kyun 60 P-006 Park, Eun Jae 72,75,88,89 P-055,P-066, P-067,P-068, P-117,P-121 Park, Sang Min 104 P-181 Park, Se-Ho 66,77 P-030,P-074 Park, Eunjin 76 P-071 Park, Seok-Rae 80,111 P-085,P-210 Park, Ga Young 86 P-112 Park, Seong Jeong 97 P-155 Park, Gayoung 73,94,113 P-057,P-144, P-218 Park, Seong-Hwan 101 P-169 Park, Shin Ae 107 P-195 Park, Hae-Ran 87 P-114 Park, So Young 101 P-170,P-171 Park, Hanul 114 P-222 Park, Sol-Ji 61 P-010 Park, Heon Woo 106 P-189 Park, Soojin 74 P-063 Park, HongJai 69 P-041 Park, Su-Ho 67 P-034 Park, Hyo Jin 54,81 OP 2,P-091 Park, Sun 81 P-089 Park, Hyun Tae 66 P-032 Park, Sung-Gyoo 68,74 P-038,P-062 Park, Hyun-Kyu 110 P-205 Park, Sungha 90 P-126 Park, Ji-Hwan 67 P-035 Park, Sung-Hwan 78 P-079 Park, Jiyun 114 P-222 Park, Sung-Moo 87,110 P-116,P-207 132 The 2014 Fall Conference of the Korean Association of Immunologists The 2014 Fall Conference of the Korean Association of Immunologists Author Index Author Page Session Park, Yeong Min 104,108 P-184,P-200 Park, Yeong-Min 109 Park, You Soo Page Session Sato, Noriko 114 P-223 P-202 Seo, Goo-Young 79,99 P-083,P-164 101 P-170,P-171 Seo, Hyungseok 102 P-176 Park, Young Chul 69,70 P-042,P-046 Seo, Jin-Won 92 P-133 Park, Young-Jun 102 P-176 Seo, Kwang-Won 62 P-016 Park, Youngjun 63 P-018 Seo, Kyoung Yul 99,100,110 P-163,P-168, P-206 Park, Yunji 66,97 P-031,P-155 Seo, Sang-Hwan 111 P-209 Poudel, Barun 112 P-215 Seo, YB 107 P-194 Pyo, Minji 72,75,88,89 P-055,P-066, P-067,P-068, P-117,P-121 Seol, Min A 69,105 P-044,P-188 Seong, Seung-Yong 82,84 P-094,P-102 Sherman, Linda 6 PL II Shim, Aeri 68,86 P-038,P-110 Shim, Byung Shik 110 P-207 R Author Ra, Eun A. 94 P-141 Rah, Hyung Chul 102 P-174 Rhee, Joon Haeng 40,107 BS VI-2,P-196 Shim, Da Hee 85 P-108 Rhie, Gi-eun 89,91,109 P-122,P-123, P-131,P-201 Shim, Do-Wan 77,79 P-075,P-081 Rho, Sangchul 67 P-035 Shim, Insop 77 P-073 Rho, Semi 96 P-152 Shim, Seung Hyun 96 P-152 Rivollier, Americ 10 SL I Shin, Dasom 74 P-063 Rogers, John A. 113 P-218 Shin, Eui-Cheol 50,53,59,62,65, 66,76 Roh, Kug Hwan 61,106 P-010,P-191 BS VIII-4,OP 1, P-001,P-013, P-028,P-029, P-072 Rudra, Dipayan 39 BS VI-1 Shin, Hyun Seok 94 P-144 Ryu, Hwa Sun 72,75,88,89 P-055,P-066, P-067,P-068, P-117,P-121 Shin, Jeon-Soo 72 P-056 Shin, Ji-Hyeon 80,81 P-086,P-092 Shin, Jung Hoon 66,77 P-030,P-074 Shin, Jung U 74 P-061 Shin, Jun-Seop 79 P-084 Shin, Sung-Jae 93,108 P-138,P-197 Shin, Tae-Hoon 62 P-016 Shin, Woo-Young 77 P-075 Sim, Ji Hyun 69,105 P-044,P-188 Smith, Trevor 6 PL II Ryu, Jae-Sook 87 P-115 Ryu, Ji Hyeong 106 P-189 Ryu, Ji In 107,108 P-195,P-197 Ryu, Jung-Hwa 100 P-166 Ryu, Kyung-Ha 100 P-166 S Saitoh, Tatsuya 37 BS V-3 Sakaguchi, Shimon 12 SL II Sanchez, Melissa D. 55,90 OP 3,P-127 The 2014 Fall Conference of the Korean Association of Immunologists 133 The 2014 Fall Conference of the Korean Association of Immunologists Author Author Index Page Session Page Session Sohn, Dae-Hee 59 P-002 T Sohn, Hyun-Jung 59 P-002 Tran, Vuvi G. 88 P-120 Son, Woo-Chan 110 P-205 U Song, Boyeong 102 P-176 Uhm, Jin Bum 72 P-055 Song, Dae-Sub 93 P-139 Upadhyay, Vaibhav 65 P-025 Song, Eunjoo 56,113 OP 4,P-219 V Song, Hyun Keun 106 P-191 Verdeil, Grégory 6 PL II Song, Hyun-Keun 61 P-010 Verma, Ravi 112 P-213 Song, Hyung Geun 102 P-174 Verma, Vivek 107 P-196 Song, Hyun-Ouk 87 P-115 W Song, Jae-Hyoung 86 P-110 Wi, Young Jin 66 P-032 Song, Jie-Young 76,80 P-071,P-088 Woo, Dong-Cheol 102 P-173 Song, Jiseon 71 P-052 Woo, So-Youn 100 P-166 Song, Kyung-Hee 80 P-088 Wui, Seo Ri 107,108 P-195,P-197, P-198 Song, Man Ki 87,107,110 P-116,P-195, P-207 X Song, Min-Suk 53,66 OP 1,P-029 Xue, Qianfei 62 P-014,P-015 Song, Mi-Young 62,97 P-013,P-155 Y Song, Sukgil 63 P-018 Yan, Guang Hai 62 P-014,P-015 Song, Sukhyun 56,113 OP 4,P-219 Yang, Bo-Gie 29 BS III-3 Song, You Chan 104 P-182 Yang, Chul-Su 85 P-105 Song, Youkyong 90 P-125 Yang, Eun-Jeong 92 P-133 Song, Young Jo 109 P-204 Yang, Hyun Gul 112 P-214 Song, Youyol 102 P-173 Yang, Jae-Wook 61 P-010 Suh, Suk Hyo 85 P-108 Yang, Jeonga 111 P-211 Sun, Xiao 77 P-075 Yang, Kwangmo 101 P-170,P-171 Sung, Pil Soo 59 P-001 Yang, Yoon-Sun 68 P-040 Sung, YC 107 P-194 Ye, Minsook 77 P-073 Sung, Young Chul 97,107,111,112 P-155,P-196, P-211,P-214 Yeo, Seung Geun 82 P-095 Yeom, Junseok 111 P-211 Sung, Young-Chul 62 P-013 Yeom, Min-Joo 93 P-139 Surh, CD 107 P-194 Yeon, Seung-min 63 P-020 Surh, Charles D. 66,70,100 P-031,P-048, P-165 Yi, Eugene C. 61 P-009 134 Author The 2014 Fall Conference of the Korean Association of Immunologists The 2014 Fall Conference of the Korean Association of Immunologists Author Index Author Page Session Yi, Jae Hyuk 102 P-174 Yi, Young-Joo 61,77 P-012,P-076 Yoo, Seung-Ah 67 P-035 Yoo, Su-Jin 60 P-008 Yoo, YoungSik 99 P-161 Yoo, Young-Sik 99 P-162 Yoo, Yung Choon 111 P-210 Yoo, Yung-Choon 80 P-085 Yoon, Bo Ruem 60 P-008 Yoon, Gun Young 104 P-184 Yoon, Hee-Kyung 80 Yoon, Hye-Kyung Author Page Session Youn, Hyewon 21 BS I-3 Youn, Jeehee 64,71 P-023,P-024, P-049 Youn, Je-In 84 P-102 Yu, Hak Sun 60 P-006 Yu, Hyeong Gon 72 P-053 Yu, Qing 97 P-156 Yuk, Jae-Min 91 P-129 Yun, Chae-Ok 106 P-190 Yun, Cheol-Heui 87,98,110 P-116,P-158, P-207 P-085 Yun, Jun-Won 62 P-016 71,82 P-050,P-094 Yun, Su-jin 81 P-089 Yoon, Jihye 85 P-108 Z Yoon, Joo Chun 90 P-125 Zhang, Wei 97 P-156 Yoon, Sang Soon 102 P-174 Zhou, Jie H.S. 14 SL III Yoon, Sangchul 100 P-168 ㄱ Yoon, Sun-Woo 111 P-209 권정근 118 LS IV You, Sungyong 73 P-059 ㅅ Youm, Ji-Young 106 P-192 사승환 118 LS III The 2014 Fall Conference of the Korean Association of Immunologists 135 Executive Committee President In-Hong Choi Yonsei University Vice President Jong-Seok Lim Sookmyung Women's University Vice President-Elect Yeong Min Park Konkok University Secretary General Man Ki Song International Vaccine Institute Vice Secretary General Sang-Bae Han Chungbuk National University Jeon Soo Shin Yonsei University Kyung-Mi Lee Korea University Chong-Kil Lee Chungbuk National University Kyungjae Kim Sahmyook University Eui-Cheol Shin Korea Advanced Institute of Science and Technology Seong Kug Eo Chonbuk National University Ki-Young Lee Sungkyunkwan University Eun Young Choi Seoul National University Myung-Shin Jeon Inha University Wan-Uk Kim The Catholic University Seok Chan Eun Seoul National University Bundang Hospital Jong Hwan Park Konyang University Seok-Rae Park Konyang University Sung Jae Shin Yonsei University Je-Wook Yu Yonsei University Kee-Jong Hong Korea National Institute of Health Heung Kyu Lee Korea Advanced Institute of Science and Technology Eui-Cheol Shin Korea Advanced Institute of Science and Technology Sang-Jun Ha Yonsei University Kyoungho Suk Kyungpook National University Je-Wook Yu Yonsei University Scientific Committee Editorial Committee Finance Committee Planning Committee Information Committee International Cooperation Committee Auditor Scientific Committee Director Jeon Soo Shin Yonsei University Vice Director Kyung-Mi Lee Korea University Dae-Ki Kim Chonbuk National University Hyoung-Pyo Kim Yonsei University Chang-Hwa Song Chungnam National University Je-Wook Yu Yonsei University Seung-Hyo Lee Korea Advanced Institute of Science and Technology Won-Woo Lee Seoul National University Myoungho Jang POSTECH Yong Woo Jung Korea University Youngnim Choi Seoul National University Je-Min Choi Hanyang University Kee-Jong Hong Korea National Institute of Health Member The 2014 Fall Conference of the Korean Association of Immunologists Publisher Editor Published by In-Hong Choi Jeon Soo Shin Printed by The Korean Association of Immunologists Rm. 701, The Korea Science & Technology Center, 22, Teheran-ro 7-gil, Gangnam-gu, Seoul 135-703, Korea Tel: 82-2-797-0975, Fax: 82-2-797-0976, Homepage: www.ksimm.or.kr, www.immunenetwork.org E-mail: [email protected] Published on November 6, 2014 MEDrang Inc. Tel: 82-2-325-2093, Fax: 82-2-325-2095 E-mail: [email protected] Homepage: www.medrang.co.kr 2014년도 대한면역학회 추계학술대회 개최를 위한 협력회사의 성원에 감사드립니다. 고마바이오텍(주) 국제백신연구소 머크(주) 머크 밀리포아 사업부, 바이오사이언스 목암연구소 밀테니바이오텍 바이오클론(주) 벡톤디킨슨코리아(주) 셀트리온제약 신한금융피비기업센터 에이비온바이오메드 에펜도르프코리아(주) 엘케이바이오사이언스 인하대병원 국가지정 소화기질환 의료제품 유효성평가 서비스센터 제넥신 종근당 (주)녹십자 (주)웅비메디텍 (주)코사바이오 (주)코아텍 (주)킴앤프렌즈 (주)태산메디칼 (주)페프로텍코리아 (주)한국얀센 진원생명과학(주) 코람바이오텍 하나바이오텍 한국노바티스 한국로슈 한국먼디파마 한국바이오래드 한국벡크만쿨터(주) 한국아스텔라스제약 한국애브비(주) 한국화이자제약 한화제약(주)
