Download JSPS 1st seminar abstracts

JSPS Asian Core Program 1st annual seminar abstract 1-1
Prof. Kenzaburo Tani
Hematopoietic Cell Differentiation from Common Marmoset (Callithrix jacchus)
Embryonic Stem Cells by Their Genetic Manipulation Using the Third
Generation Lentiviral Vector.
Ryo Kurita 1, Erika Sasaki 2, Tomoko Yokoo 1, Kashiya Takasugi1, Yan Dong4,
Hideyuki Imoto1, Youko Suehiro1, Yoshimiti Tachikawa1,Takashi Hiroyama 3,
Yukoh Nakazaki 1, Kiyoko Izawa 4, Hajime Ishii 2, Yoshikuni Tanioka 2,
Kisaburo Hanazawa 5, Yuan Son Bai 4, Yasushi Soda 4 and Kenzaburo Tani 1.
1 Molecular Genetics, Bioregulation, Fukuoka, Fukuoka, Japan, 812-8582; 2
Division of Animal Experimentation, Central Institute of Experimental
Animals, Kawasaki, Kanagawa, Japan, 216-0001; 3 Cell Engineering Division,
Riken Bio Resource Center, Cell Bank, Tukuba, Ibaragi, Japan, 305-0074; 4
Division of Molecular Therapy, Institute of Medical Science, University of
Tokyo, Tokyo, Tokyo, Japan, 108-8639 and 5 Department of Urology, Juntendo
University School of Medicine, Tokyo, Japan.
We recently demonstrated that non-human primates of common marmosets (CM)
are suitable for the preclinical studies of hematopoietic stem cell therapy
and established CM ES cell lines (Sasaki, E. et al., Stem Cells 2005). In
this study, we investigated the in vitro and in vivo differentiation of CM
ES cells to hematopoietic cells by exogenous gene transfer methods. The
results demonstrated the feasibility of gene modified ES cell therapy.
First, we tested various in vitro culture conditions including embryoid body
formation or co-culturing with stromal cells to induce hematopoietic cells,
but the hematopoietic activities were rather low. The expression of CD45 and
gata1 could not be detected in both conditions, suggesting that the culture
conditions were lacking essential factor for induction of hematopoietic
cells from CM ES cells. Next we examined gene transduction methods by using
VSV-G pseudotyped human immunodeficiency virus (HIV) vectors which were
constructed to express hematopoietic genes such as marmoset tal1/scl, gata1,
gata2, Lh2 and human hoxB4 genes under the EF1 promoter and transduced them
into CM ES cells. Only when tal1/scl gene was transduced, hematopoietic
induction from CM ES cells was dramatically stimulated and multi-lineage
blood cells consisting of erythroid cells, granulocytes, macrophages and
megakaryocytes, were observed by immunochemical and morphological analyses.
JSPS Asian Core Program 1st annual seminar abstract 1-2
Prof. Kenzaburo Tani
Furthermore, FACS analysis and RT-PCR results showed that CM CD34, one of
the hematopoietic stem cell markers, was expressed in the tal1/scl
overexpressed-EB cells. These results were similarly obtained from
experiments using three independent CM ES lines. After the
xenotransplantation of ES-derived cells into immunodeficient mice, CM CD45+
cells and immature erythroid cells were observed in the ES-tal1-injected
mice, indicating that enforced expression of tal1/scl into ES cells led to
highly efficient hematopoietic cell differentiation both in vitro and in
vivo. Above results suggested that the transduction of exogenous tal1/scl
cDNA into ES cells by HIV vector was the promising method for the efficient
differentiation from CM ES cells to hematopoietic stem cells. Determination
of both long-term hematopoietic reconstitution capacity and safer
intracellular delivery of tal1/scl gene products are under investigation for
the purpose of developing new hematopoietic stem cell therapy.
JSPS Asian Core Program 1st annual seminar abstract 2
Dr. Kennosuke Karube
The pathology of adult T-cell lymphoma/leukemia
Kennosuke Karube, Koichi Ohshima
Department of Pathology, Kurume University
Adult T cell lymphoma/leukemia (ATLL) is caused by human T cell
lymphotropic virus type I (HTLV-1) and characterized by aggressive clinical
course and poor prognosis. ATLL is prevalent in areas endemic for HTLV-1,
including Kyushu, which locates southwestern Japan, and the Caribbean
basin. Until now, many studies about HTLV-1 virus and the mechanism of
infection progressed, but the prognosis of ATLL is still poor and the
oncogenic mechanism of ATLL has not been known completely.
In this seminar, we would like to introduce our study analyzing the
morphological,
epidemiological,
genetical,
and
immunohistochemical
features of ATLL.
(1)
The incidence of ATLL in Japan
(2)
The morphological feature of ATLL
(3)
The immunohistochemical feature of ATLL
(4)
The association of ATLL lymphoma cells and CD4+CD25+
regulatory T cells: FoxP3 expression in ATLL and its effect on the clinical
feature
JSPS Asian Core Program 1st annual seminar abstract 3
Prof. Naotaka Hamasaki
Japanese Thrombophilia
Protein S and Protein C gene mutations in Japanese deep vein thrombosis patients.
Naotaka Hamasaki, M.D., Ph.D.
Faculty of Pharmaceutical Science
Nagasaki International University
Coagulation factor V Leiden has not been detected in Japanese patients
suffering from thrombosis. Hitherto the constitutional background of Japanese
thrombotic patients has never been systematically examined. We have performed a
systematic investigation to determine pathogenesis for deep vein thrombosis in
Japanese population. Routine coagulation and fibrinolysis tests were performed to
determine the activities of protein S, protein C, antithrombin, plasminogen and
fibrinogen. Gene analysis was performed in thrombotic patients having the low
activities of these factors.
Our study indicates that the frequency (19/85=0.22) of mutations of protein S
gene in the Japanese patients was 5-10 times higher than that of mutations of protein S
gene in Caucasian patients, and the frequency (8/85=0.09) of mutations of protein C
gene was almost three times higher than that of Caucasian patients. Protein S
reinforces the activated protein C activity and cooperatively regulates the coagulation
activity with the activated protein C. Since factor V Leiden shows resistance to the
protein S/protein C system, coagulation regulation by the protein S/protein C system
profoundly fails in individuals having factor V Leiden, yielding the same phenotypes
as those with reduced protein S/protein C anticoagulation activity. It is interesting to
note that the frequency of factor V Leiden (20-50%) in Caucasian deep vein
thrombosis patients is similar to that (30%) of heterozygous gene mutations of the
protein S/protein C anticoagulation system in Japanese deep vein thrombosis patients.
Key Words: Deep vein thrombosis; protein S gene mutation; protein C gene
mutation; factor V Leiden; Japanese thrombophilia.
JSPS Asian Core Program 1st annual seminar abstract 4
Prof. Ampaiwan Chuansumrit
THROMBOEMBOLISM IN THAI CHILDREN
Ampaiwan Chuansumrit, M.D., Nongnuch Sirachainan, M.D.
Department of Pediatrics, Faculty of Medicine, Ramathibodi Hospital, Mahidol
University, Bangkok, Thailand
Objective:
To analyze the etiology, management and outcome of Thai pediatric
patients with thromboembolism.
Methods: A retrospective study of patients admitted at the Department of Pediatric,
Ramathibodi Hospital from 1987 to 2003, was conducted.
Results: Total 108 patients with thromboembolisms were enrolled in the study. The
sites of thromboembolism were in the venous sinus or arteries of the central nervous
system 33%; the skin with necrosis, 20%; deep vein of the extremities, 16%; the toes
and fingers with gangrene, 14%; and other sites such as the heart and lugs, 17%.
Most of the patients had triggering conditions (54%) including endothelial injury,
stasis of blood and alteration of coagulation factors.
Factor V Leiden and
prothrombin 20210 were not found among the studied patients; however, low levels of
antithrombin III, protein C or protein S were found in 39% (22/57) of the patients.
Most of these were transient except for three who were diagnosed with homozygous
(n=3) and heterozygous (n=4) protein C deficiency.
The management included
administration of standard heparin or low molecular weight heparin, if not
contraindicated replacement of fresh frozen plasma 10 ml/kg twice a day and
treatment of triggering and underlying conditions.
The fatality rate was 14%.
Subsequent episodes of thromboembolism were found in 13 patients who did not
receive appropriate management.
Conclusion: Comprehensive investigation and specific treatment for pediatric patients
are emphasized in order to prevent recurring thromboembolic episodes.
JSPS Asian Core Program 1st annual seminar abstract 5
Assoc.Prof. Shuji Shimizu
High-quality interactive telecommunication system using super-fast Internet
connections and its application to hematology
Kyushu University, Fukuoka, Japan
Shuji Shimizu, Naoki Nakashima, Nobuhiro Torata, Koji Okamura, Koichiro Muta,
Masao Tanaka
Amid rapid change in medical knowledge and patient care, education and training
for younger generation is of utmost importance beyond geographic borders.
International telecommunication system via super-fast broadband Internet, which
was established between Japan and Korea in 2003 and can transmit moving images
without any loss of quality, was reported to be a powerful tool for this purpose. The
aim of our study was to expand our advanced system to whole range of Asia-Pacific
regions and to widen our application fields over rapidly growing optic fiber network.
Kyushu University Hospital in Japan was newly linked to over 20 medical
institutions or meeting venues in China, Taiwan, Thailand, Vietnam, Singapore, and
Australia through domestic and international research and development networks.
A digital video transport system (DVTS), which can transform digital video signals
directly to Internet protocol with the bandwidth of 30 Mbps per line, and cipher
security programs to protect patient’s privacy were used.
Of the 60 international teleconferences conducted during three years, real-time
demonstrations and interactive teleconferences with recorded videos were included.
Not only two stations were connected pier-to-pier, but also multiple stations were
connected without any conversion to analog signals. The medical fields involved
were surgery, endoscopy, hematology, medical informatics, nursing, health care, etc.
The network remained stable, and the time delay between stations was restricted to
0.3 – 1.0 sec.
We have established a high-quality telecommunication system in wide area of
Asia-Pacific, and this is the first time to apply high-speed Internet technology to
medicine in such a big international scale. Because it is not only high-quality but
also economical and easily accessible, we believe this system will promote worldwide
expansion of efficient medical education and active academic exchanges.
JSPS Asian Core Program 1st annual seminar abstract 6
Dr. Nopphol Pausawasdi
Telemedicine: Siriraj’s education and research network
Nopphol Pausawasdi, PhD.
Faculty of Medicine Siriraj Hospital, Mahidol University
The Faculty of Medicine Siriraj Hospital has three major missions which are (a)
provides medical education (b) produces medical research and (c) provides medical
care. To accomplish those missions effectively, the faculty has consider advanced
technology which would enhance teaching and learning activities, research projects
and collaborations, and medical care skills and knowledge. Telemedicine is a
technology that helps the faculty to establish collaborations with other medical
institutions over the world which allows us to create medical education and research
network. In 2004, the faculty initially utilized telemedicine in a Siriraj annual
medical conference to allow medical staffs who were working in Siriraj hospital and
other private hospitals in Bangkok to see presentations from the conference venue as
well as to be able to communicate with speakers. In the same year, the faculty began
to provide medical education to medical students who studied in Ratchaburi hospital
which is located in Ratchaburi province. Moreover, we also collaborated with a group
of doctors from Kyushu University Hospital to conduct a telemedicine session from
the Faculty of Medicine Siriraj Hospital to Kyushu University Hospital. Since then
the Faculty of Medicine Siriraj Hospital set up its telemedicine internet network
connection with great supports from many national and international organizations
including Mahidol University, Uninet, Nectec, and Kyushu University Hospital.
Currently the faculty is able to utilize telemedicine technology in everyday routine
working basis and other special events. We collaborate with many local and global
medical institutions including Ramathobodi Hospital, Ratchaburi Hospital,
Samitivet Hospital, Bangkok General Hospital, and those of APAN medical working
group such as Kyushu University Hospital and Seoul National University Bundang
Hospital in order to develop education and research network among doctors and
medical staffs around the world.
The Faculty of Medicine Siriraj Hospital will
continue to expand its network to other medical institutions to enrich medical
information exchange which would help developing medical education and research
of the faculty and the others.
JSPS Asian Core Program 1st annual seminar abstract 7-1
Prof. Shinji Nakao
Specific antibodies to moesin,
a membrane-cytoskeleton linker protein, in the serum of
patients with aplastic anemia: their incidence and possible role in the pathophysiology of
bone marrow failure.
Shinji Nakao, Hiroyuki Takamatsu.
Cellular Transplantation Biology, Kanazawa University
Graduate School of Medical Science
Although aplastic anemia (AA) is a T-cell mediated disease, recent studies have revealed the
presence of antibodies (Abs) specific to proteins derived from hematopoietic progenitor cells in
the serum of AA patients.
pathophysiology of AA.
It is as yet unclear whether these auto-Abs play some roles in the
We recently demonstrated that Abs specific to moesin, a
membrane-cytoskeleton linker protein in the cytoplasm, were detectable in approximately 37%
of AA patients.
Some reports identified moesin-like molecules on the surface of blood cells
such as T cells and macrophages.
It is therefore conceivable that anti-moesin Ab in AA patients
may bind these immune cells and modulate hematopoietic function of AA patients.
To test
these hypotheses, we first studied the expression of moesin on various types of blood cells using
monoclonal Ab specific to moesin (clone 38/87). Flow cytometry detected the expression of the
protein recognized by anti-moesin Ab on T cells and monocytes from healthy individuals, acute
monocytic leukemia cells lines including U937 and THP-1, and an acute T-lymphoblastic
leukemia cell line, Molt-4, but failed to detect the molecule on CD34 + cells from healthy
individuals and myeloid leukemia cell lines as well as B-lymphoblastic leukemia cell lines.
Treatment of THP-1 cells with phorbol 12-myristate 13-acetate (PMA)/lipopolysaccharide (LPS)
augmented the expression level of moesin.
To confirm the expression of the moesin-like
protein on the surface of monocytic leukemia cell lines, Molt-4 and THP-1 were treated with
sulfo-NHS-SS-biotin, and the cell surface proteins were isolated with avidin-fixed column, and
were subjected to Western blotting and peptide mass fingerprinting.
Western blotting with
anti-moesin monoclonal Abs showed two clear bands of proteins (75 kD and 80 kD); an amino
acid sequence compatible with moesin was confirmed in the protein eluted from the 80 kD band.
Next, we purified anti-moesin Abs from AA patients’ sera using affinity chromatography with
recombinant moesin protein.
Western blotting showed binding of the serum-derived Abs to a
fraction of surface proteins of Molt-4, U937 and THP-1.
When THP-1 cells were incubated in
the presence of PMA and LPS with 5 g/ml of control IgG or anti-moesin Abs derived from an
AA patient’s serum, TNF- production from THP-1 cells stimulated by anti-moesin Abs was
1.9-2.3 times as much as that from the control culture depending on the concentration of LPS.
Incubation of THP-1 cells in the presence of monoclonal anti-moesin Abs showed the similar
augmentation of TNF- production.
These results indicate that anti-moesin Abs may be
JSPS Asian Core Program 1st annual seminar abstract 7-2
Prof. Shinji Nakao
involved in the suppression of hematopoiesis of AA patients by stimulating TNF- production
from monocytes.
JSPS Asian Core Program 1st annual seminar abstract 8
Dr. Yukimi Sakoda
Pathophysiology and treatment of chronic GVHD after allogeneic hematopoietic
stem cell transplantation
Sakoda Yukimi
Department of Medicine and Biosystemic Science, Kyushu University
Chronic graft-versus-host disease (GVHD) is the most common cause for poor
long-term outcomes after allogeneic bone marrow transplantation (BMT).
Unfortunately, despite success in the prevention of acute GVHD by the introduction
of cyclosporine, the incidence of chronic GVHD has not decreased. However, the
pathophysiology of chronic GVHD still remains poorly understood. Recent animal
studies suggested that donor CD4+ T cells play a critical role in the induction of
chronic GVHD, thus T cell-depletion (TCD) could reduce chronic GVHD as well as
acute GVHD. However, the first randomized study comparing TCD-BMT with T
cell replete BMT found that TCD reduces rates of acute GVHD, but not chronic
GVHD, thus suggesting that thymic-dependent T cells play a role in mediating
chronic GVHD. We tested the hypothesis that the impaired thymic function of the
recipients will permit the emergence of pathogenic T cells that cause chronic GVHD
using chimeric mice where thymic negative selection was impaired. Lethally
irradiated C3H/HeN (H-2k) recipients were reconstituted with TCD bone marrow
cells from MHC class II-deficient B6 (H-2b) mice. These mice developed diseases
that showed all of the clinical and histopathological features of human chronic
GVHD. Thymectomy prevented chronic GVHD, thus confirming the causal
association of the thymus. We also found that CD4+ T cells isolated from chronic
GVHD mice were primarily donor-reactive and adoptive transfer of these CD4+ T
cells caused chronic GVHD in recipient mice in the presence of donor-derived
antigen presenting cells. Our results demonstrate for the first time that T cells which
escape from negative thymic selection could cause chronic GVHD. These results
also suggest that self reactivity of donor T cells play a role in this chronic GVHD
and improvement in the thymic function may have a potential to decrease chronic
GVHD. A recent study from another group showed donor B cells play a role in
chronic GVHD. We therefore conduct a phase II trial of anti-B cell therapy with
rituximab in steroid refractory chronic GVHD.