Download ARVO 2012 Annual Meeting Abstracts

ARVO 2012 Annual Meeting Abstracts
530 Autoimmune Ocular Disease
Thursday, May 10, 2012, 8:30 AM - 10:15 AM
Hall B/C Poster Session
Program #/Board # Range: 6228-6256/D1088-D1116
Organizing Section: Immunology/Microbiology
Contributing Section(s): Cornea,Retina+,Retinal Cell Biology
Program Number: 6228 Poster Board Number: D1088
Presentation Time: 8:30 AM - 10:15 AM
Intraocular Th1/Th17 Cells Coexpressing IL-10: Tregs that Prevent
Recurrent EAU?
Ulrike Kaufmann, Maria Diedrichs-Moehring, Gerhild Wildner. Section of
Immunobiology, Ophthalmology, Clinic of the Ludwig-Maximilians-University,
Munich, Germany.
Purpose: The strict compartmentation of T cells in Th1, Th2 and Th17 was
mitigated with the discovery of T cells that simultaneously produce IFN-γ and IL17. They are found in chronically inflamed tissues and are not stable with respect to
their cytokine expression. The role of these cells is still unknown. We have
investigated intraocular T cells during the course of EAU with respect to their
cytokine and Foxp3 expression in our rat model of monophasic and relapsingremitting uveitis.
Methods: After induction of EAU with PDSAg (monophasic EAU) or R14
(relapsing EAU) in CFA we collected intraocular cells at various time points during
ocular inflammation and stained them ex vivo for TCRαβ and intracellular IFN-γ,
IL-17, IL-10 and Foxp3 expression.
Results: During the course of PDSAg-induced, monophasic uveitis intraocular T
cell populations coexpressing IFN-γ and IL-17 increased from onset via peak to
remission of EAU. IL-10+ and IL-17+/IL-10+ cells displayed a similar population
dynamic, the latter increased to about 30% of the Th17 population at remission.
Like Foxp3+ cells, IFN-γ+/IL-10+ cells were a minor population remaining stable
during the course of monophasic EAU. In R14-induced relapsing EAU the number
of IL-17+ and IFN-γ+/IL-17+ cells decreased at remission, while IFN-γ+ cell
numbers remained elevated. IL-17+/IL-10+ cells decreased during remission of
R14-induced EAU, while IFN-γ+/IL-10+ and Foxp3+ cells were also minor
intraocular lymphocyte populations.
Conclusions: The change of the intraocular T cell populations during EAU, the
large numbers of T cells producing multiple cytokines and the differences observed
between monophasic and relapsing disease points to a strong population dynamics
in the eye, potentially influencing the course of EAU. The strong increase of IL10+ T cells in the eyes during monophasic EAU suggests a regulatory role of these
cells with a potential to prevent relapses. Foxp3+ cells seem to play no role in
preventing recurrent EAU.
Commercial Relationships: Ulrike Kaufmann, None; Maria DiedrichsMoehring, None; Gerhild Wildner, None
Support: Deutsche Forschungsgemeinschaft SFB 571
Program Number: 6229 Poster Board Number: D1089
Presentation Time: 8:30 AM - 10:15 AM
Cd4+ Foxp3+ Cd25Bright T Regulatory Cells Population In Ocular
Sarcoidosis
Alexis Pinel1A, Alexis Mathian1B,2, Makoto Miyara1B, Catherine CHAPELONABRIC1B, Christophe Parizot1C, Du Boutin1B, Zahir Amoura1B, Guy Gorochov1C,
Phuc Lehoang1A, Bahram Bodaghi1A,3. AOphthalmology, BInternal medecine,
C
Immunology, 1CHU Pitie-Salpetriere, Paris, France; 2INSERM UMR-S 945, Paris,
France; 3U972 INSERM, Paris, France.
Purpose: CD4+ FOXP3 CD25bright T regulatory cells (Treg) are expanded in
systemic sarcoidosis wich is a chronic inflammatory disorder characterized by
noncaseating granulomas. The effector Treg expansion is already described in
pulmonary and renal sarcoidosis. The main objective of this study is to describe the
circulating Treg population (naive and effector Treg) in uveitis and in particular in
ocular sarcoidosis.
Methods: Patients followed for uveitis were included. Clinical, biological and
pathological features were analyzed. The patients were divided in 4 groups :
definite ocular sarcoidosis among the international criteria for the diagnosis of
ocular sarcoidosis from the International Workshop on Ocular Sarcoidosis (IWOS),
suspected ocular sarcoidosis but not biopsy proved, idiopathic uveitis and uveitis of
other origin. Treg populations were measured in blood using fluocytometry
analysis.
Results: 103 patients were included with 19 definite ocular sarcoidosis, 12
suspected sarcoidosis, 41 idiopathic uveitis and 31 uveitis of other diagnosis. In the
definite ocular sarcoidosis and the suspected sarcoidoisis groups, the median
effector Treg rate was 2,54 and 2,7 which was significantly higher than in the
control group (p<0,0001). There were no statistical difference between the Treg
effector rate of idiopathic uveitis (0,86), uveitis of other diagnosis (0,73) and the
control group. In the definite ocular sarcoidosis group, the Treg was significantly
higher compared to idiopathic uveitis (p=0,046) and to uveitis of other origin
(p=0,009). The naive Treg population showed no statistical difference between the
4 groups.
Conclusions: Definite ocular sarcoidosis and suspected ocular sarcoidosis are
associated with peripheral effector Treg expansion. There is no alteration of Treg
population in idiopathic uveitis and uveitis of other origin. The naive Treg
population is not expanded in the 4 groups. This test could be used as a new
diagnostic tool in uveitis in particular when sarcoidosis is suspected.
Commercial Relationships: Alexis Pinel, None; Alexis Mathian, None; Makoto
Miyara, None; Catherine Chapelon-abric, None; Christophe Parizot, None; Du
Boutin, None; Zahir Amoura, None; Guy Gorochov, None; Phuc Lehoang,
None; Bahram Bodaghi, None
Support: None
Program Number: 6230 Poster Board Number: D1090
Presentation Time: 8:30 AM - 10:15 AM
Assessment of Th1, Th2, and Th17 Cells in Birdshot Retinochoroidopathy
Paul Yang, C. S. Foster. Ophthalmology, Massachusettes Eye Research and
Surgery Institute, Cambridge, MA.
Purpose: To correlate the serum levels of 20 cytokines from the T helper (Th)1,
Th2, and Th17 pathways with disease activity in birdshot retinochoroidopathy
(BSRC).
Methods: Single center cohort study of 4 BSRC patients, and 4 normal controls.
Immunomodulatory therapy (IMT) and disease status were recorded for each
patient. Disease activity was assessed with visual acuity, visual field, fundus exam,
optical coherence tomography, fluorescein angiography, and electroretinography.
Sera were collected, and a quantitative multiplex sandwich ELISA-based
microarray assay was used to quantify 20 cytokines: interleukin (IL)-1beta, IL-2,
IL-4, IL-5, IL-6, IL-10, IL-12p70, IL-13, IL-17, IL-17F, IL-21, IL-22, IL-23, IL28, interferon (IFN)-gamma, macrophage inflammatory protein (MIP)-3alpha,
transforming growth factor (TGF)-beta1, tumor necrosis factor (TNF)-alpha, TNFbeta, and granulocyte macrophage colony-stimulating factor (GM-CSF). Cytokine
levels from patients were compared with normal controls and correlated with IMT
and disease activity.
Results: IL-21 (p = 0.01), IL-23 (p = 0.02), TGF-beta1 (p = 0.02), and TNF-beta (p
= 0.02) were significantly elevated in the sera of BSRC patients compared to
normal controls. IL-17 (p = 0.31) and IFN-gamma (p = 0.85) serum levels were not
significantly elevated. The subject that had been in IMT-dependent clinical
remission for 2 years, had the lowest levels of IL-21 and IL-23 levels. IL-21 and
IL-23 levels had a correlation coefficient of r = 0.93.
Conclusions: Th17 cells play an important role in autoimmune disease and uveitis.
Although standard ELISA assay has not shown elevations of IL-17 in the sera of
BSRC patients, a recent multiplex immunoassay of 23 cytokines in BSRC patients
revealed that IL-17 and proinflammatory cytokines were concentrated in the
aqueous fluid. We confirm that IL-17 is not elevated in the sera of BSRC patients,
however both IL-21 and IL-23 levels were significantly elevated and correlated,
suggesting that peripheral cytokines may promote Th17 cells in an organ specific
manner. In addition, higher IL-21 and IL-23 levels may correlate with active
disease.
Commercial Relationships: Paul Yang, None; C. S. Foster, None
Support: Ocular Immunology and Uveitis Foundation
Program Number: 6231 Poster Board Number: D1091
Presentation Time: 8:30 AM - 10:15 AM
Regulatory T-Cells In Peripheral Blood of Patients With Birdshot
Retinochoroidopathy
Sana S. Siddique1, Laura Amorese1, Lama Almulki2, Ana M. Suelves3, C Stephen
Foster4. 1Ophthalmology, MERSI/OIUF, Cambridge, MA; 2Ophthalmology,
Massachusetts Eye Res and Surgery Inst, Cambridge, MA; 3MERSI, Xirivella,
Valencia, Spain; 4Ophthalmology, Ocular Immunol & Uveitis Fndtn, Cambridge,
MA.
Purpose: CD4+CD25 T cells have been shown to be involved in the pathogenesis
of autoimmune diseases. We aimed to assess the percentage of T regulatory cells in
the peripheral blood of patients with active Birdshot Retinochoroidopathy (BSRC).
Methods: After obtaining informed consent, 5ml of blood was collected from a
total of twelve patients diagnosed with new onset or untreated BSRC and 5 healthy
controls. The samples were labeled and transported immediately to the Clinical
Flow Cytometry Laboratory at the Massachusetts General Hospital (MGH) for
analysis. Peripheral blood mononuclear cells were subjected to flow cytometry for
analysis of lymphocytes expressing the following cell surface glycoproteins: CD4
CD25 CD127 FOXP3. The biomarkers utilized to appropriately identify and purify
activated T-regulatory cells included CD4, CD25, CD127, Foxp3, CD3 and CD45.
Results: Samples were analyzed from 12 active BSRC patients and 5 healthy
controls. The preliminary data reveals level of activated T-cells CD4+CD25hi was
4.6% in patients and 5.1% in the controls, while the level of CD4+CD25hi
FOXP3+ regulatory T lymphocytes levels was 80.2% in patients and 78.4% in
controls.
Conclusions: Our preliminary data shows no statistical difference between the two
Copyright 2012 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2012 Annual Meeting Abstracts
groups. It is unclear whether CD4+CD25hi FOXP3+ Treg cells play a role in the
development of BSRC.
Commercial Relationships: Sana S. Siddique, None; Laura Amorese,
None; Lama Almulki, None; Ana M. Suelves, None; C Stephen Foster, None
Support: None
Program Number: 6232 Poster Board Number: D1092
Presentation Time: 8:30 AM - 10:15 AM
Clinical Course of Patients with Behcet's Uveoretinitis that Discontinued
Infliximab Therapy
Tatsushi Kawaguchi1,2, Yuko Iwasaki1,2, Sayaka Kanda2, Sunao Sugita2, Manabu
Mochizuki2. 1Ophthalmology, Tokyo Metropolitan Komagome Hospital, Tokyo,
Japan; 2Ophthalmology & Visual Science, Tokyo Medical and Dental University,
Tokyo, Japan.
Purpose: To investigate clinical course of patients with Behçet's uveoretinitis that
discontinued infliximab (IFX) therapy.
Methods: Subjects were uveitis patients associated with Behçet’s disease that
received IFX treatment at Tokyo Medical and Dental University Hospital between
2000 and 2011. Medical records of patients who discontinued IFX were reviewed
retrospectively. The patients followed less than 12 months before and after IFX
treatment were excluded. Frequency and severity of acute uveitis attacks, and
visual prognosis were evaluated.
Results: Of forty patients included in this study, nine patients (22.5%) discontinued
IFX due to adverse events. In these patients, frequency of acute uveitis attack
(mean/month) was 0.56 before IFX, 0.22 during IFX, and 0.06 after cessation of
IFX. The frequency of uveitis attacks was statistically higher before IFX compared
to during IFX (p<0.01), and after cessation of IFX (p<0.01), while there was no
statistical significance between during IFX and after cessation of IFX (p=0.12). The
severity of uveitis attacks was tend to be higher in the periods before IFX compared
to during IFX, or after cessation of IFX. The average of visual acuity was 0.25
before IFX, 0.30 during IFX, 0.37 at 12 months after cessation of IFX, and there
was no statistical significance between them.
Conclusions: Clinical course of Behçet's uveoretinitis after cessation of IFX
appears to be satisfactory with well-controlled ocular inflammation. These data
suggest a possibility for secure, planned discontinuation of IFX therapy in Behçet's
uveoretinitis.
Commercial Relationships: Tatsushi Kawaguchi, None; Yuko Iwasaki,
None; Sayaka Kanda, None; Sunao Sugita, None; Manabu Mochizuki, None
Support: None
Program Number: 6233 Poster Board Number: D1093
Presentation Time: 8:30 AM - 10:15 AM
Anti-DEC205 Mediated Delivery of Self-Antigen to Dendritic Cell Restores
Tolerance in Spontaneous EAU
Koju Kamoi1, Cristina Martin-Granados1, Corina Bobu1, Matthew E. Wikstrom2,
Mariapia A. Degli-Esposti2, Ralph M. Steinman3, John V. Forrester1.
1
Ophthalmology, University of Aberdeen, Aberdeen, United Kingdom; 2Lion Eye
Institute, University of Western Australia, Nedlands, Western Australia, Australia;
3
Rockfeller University, New York, NY.
Purpose: Dendritic cells (DCs) play critical roles in the adaptive immune response
participating both in immune activation and induction of tolerance. As an
immunotherapy, DC “vaccination” has been used in patients with cancer for
induction of immunity, but to date induction of tolerance for autoimmune disease is
still at a preclinical stage. We have previously induced tolerance to IRBP by using
antigen-pulsed immature DC and LPS-stimulated DC in a model of experimental
autoimmune uveoretintitis (EAU). Here we have used a more targeted approach to
induce tolerance by delivering antigenic peptide to different DC subsets using DCselective antibody-peptide fusion proteins in a spontaneous model of EAU.
Methods: 1.Spontaneous EAU mice was induced in B10.BR mice by crossing
single Tg 3A9 TCR mice with Tg mice expressing hen egg lysozyme (HEL) in
photoreceptor cells under control of the promoter for IRBP. 2.Anti-DEC205-HEL
fusion protein and anti-33D1-HEL fusion proteins were purified by the transfection
of expression vectors. 3. Targeting of the fusion proteins to DC after subcutaneous
injection was confirmed by immunostaining and flow cytometry of draining lymph
node cells. 4. Modulation of spontaneous EAU was evaluated clinically and
histologically. 5. Induction/expansion of regulatory T cells in eye and lymph node
after fusion protein treatment were measured by flow cytometry.
Results: Anti-DEC205-HEL and anti-33D1 fusion proteins (5ug subcut) tracked
selectively to DLN DC. Administration of anti-DEC205-HEL at d18 postnatal
prevented development of EAU, which reached maximum levels at around d 30 in
100% of untreated mice. In contrast, anti33D1-HEL led to an acceleration of EAU
development. Histological evaluation indicated that anti-DEC205-HEL markedly
reduced the numbers of infiltrating effector CD4+ T cells in the eye, while
CD4+CD25+FoxP3+ regulatory T cells in eye-draining lymph nodes were
increased by anti-DEC205-HEL, but decreased by anti-33D1-HEL.
Conclusions: Targeted delivery of antigen to specific DC subsets can promote
tolerance or immunity depending on the DC specific surface molecule. Clinical
trails in promoting immune response to tumour antigens are already in progress.
The data presented here in a preclinical model reveal the potential for such therapy
to regulate autoimmune disease.
Commercial Relationships: Koju Kamoi, None; Cristina Martin-Granados,
None; Corina Bobu, None; Matthew E. Wikstrom, None; Mariapia A. DegliEsposti, None; Ralph M. Steinman, None; John V. Forrester, None
Support: None
Program Number: 6234 Poster Board Number: D1094
Presentation Time: 8:30 AM - 10:15 AM
Monocyte-derived Macrophages in EAU Resolution
Inbal Benhar1, Anat London1, Rachel R. Caspi2, Michal Schwartz1. 1Neurobiology,
Weizmann Institute of Science, Rehovot, Israel; 2Laboratory of Immunology,
National Eye Inst/NIH, Bethesda, MD.
Purpose: We recently showed that monocyte-derived macrophages take part in
protecting retinal ganglion cells (RGCs) after glutamate-induced retinal insult, by
locally exerting an immunoregulatory phenotype. In experimental autoimmune
uveitis (EAU), macrophages are commonly associated with disease induction and
progression. Appreciating the functional diversity of monocyte-derived
macrophages, we set out to find whether the same subsets of monocyte-derived
macrophages that are involved in homeostasis restoration following glutamate
intoxication are also involved in protection against immune-mediated damage
associated with EAU.
Methods: EAU was induced in male C57BL/6J mice by injection of human
interphotoreceptor retinoid binding protein (IRBP)-derived peptide 1-20. The
infiltration of monocyte-derived macrophages to the eye along disease course was
monitored in CX3CR1-GFP bone marrow chimeric mice. Depletion experiments
were performed in order to evaluate EAU resolution in the absence of monocytederived macrophages.
Results: EAU induction resulted in the appearance of a distinct myeloid population
in the retina. Monocyte-derived macrophages infiltrated diseased retinas, and were
absent from retinas of control mice. Inhibition of this infiltration at the resolution
phase resulted in a decrease in FoxP3+ regulatory T cells, and affected the local
cytokine milieu.
Conclusions: Monocyte-derived macrophages are active cells in EAU. These cells
are present throughout disease course, affecting the local milieu in terms of
cytokines and accumulation of other immune cells, and may play a role in disease
resolution that is apparently distinct from their involvement in disease induction.
Commercial Relationships: Inbal Benhar, None; Anat London, None; Rachel
R. Caspi, None; Michal Schwartz, None
Support: None
Program Number: 6235 Poster Board Number: D1095
Presentation Time: 8:30 AM - 10:15 AM
Alpha-1 Adrenergic Stimulation Exacerbates Acute Ocular Inflammation
Through A Mechanism Mediated By Transforming Growth Factor Beta
(TGFB)
Paola A. Durand1, Yaohong Tan2, Darakshan Fatmi1, Xiaomei Xia2, Egla Suarez1,
Victor L. Perez2, Jose L. Vega1. 1Neurology, Herbert Wertheim College of
Medicine-Florida International University, Miami, FL; 2Ophthalmology, Bascom
Palmer Eye Insitute, Miller School of Medicine, Miami, FL.
Purpose: Previously we demonstrated that ocular immune privilege is abolished by
the surgical transection of ocular sympathetic nerves. Here we investigated the
effect of sympathetic adrenergic neurotransmission on acute ocular inflammation.
Methods: The selective alpha-1 adrenergic receptor (A1AR) agonist PHE (5%)
was applied every 8h for 24h to the ocular surface of mice afflicted with endotoxininduced uveitis. In some experiments topical prazosin (PRAZ), an A1AR blocker,
was applied concomitantly in order to test the specficity of A1AR-mediated effects.
Inflammation was quantified by flow cytometry of digested ocular homogenates
using antibodies against cell surface markers known to appear in the context of
acute inflammatory responses. Total aqueous humor protein quantification (via
Bradford assay), and H&E staining were also performed to further characterize
inflammation in EIU. In addition, we tested the effect of A1AR stimulation on
neutrophilic infiltration following syngeneic corneal transplantation of C57BL/6
mice, also by performing flow cytometry of corneal graft homogenates. Finally,
because A1AR stimulation increases transforming growth factor beta (TGFB), we
then tested the potential pro-inflammatory role of this TGFB increase by carrying
out EIU experiments on PHE-treated TSP-1 null mice, which exhibit a failure in
TGFB activation.
Results: A1AR stimulation with topical PHE induced dramatic exacerbations of
EIU, as reflected by flow cytometry, histology, and bradford assay results. These
exacerbated responses could be completely prevented by co-treatment with topical
PRAZ, which also significantly alleviated EIU in mice not treated with PHE.
Topical PHE also led to a significant increase in neutrophilic infiltration of
syngeneic corneal grafts. TSP-1 null mice exhibited similar EIU responses by
comparison with wildtype mice, and did not experience detectable exacerbations
after PHE treatment.
Copyright 2012 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2012 Annual Meeting Abstracts
Conclusions: Local A1AR stimulation exacerbates acute ocular inflammation via
an effect that appears to involve TGFB expression, and function.
Commercial Relationships: Paola A. Durand, None; Yaohong Tan,
None; Darakshan Fatmi, None; Xiaomei Xia, None; Egla Suarez, None; Victor
L. Perez, None; Jose L. Vega, None
Support: None
Program Number: 6236 Poster Board Number: D1096
Presentation Time: 8:30 AM - 10:15 AM
Effect Of P2Y2 Deficiency On Experimental Autoimmune Uveitis
Development
Laure E. Caspers1, Lia J. Relvas1, Remi Dewispelaere1, Maya Makhoul2, Didier
Communi2, Jean-Marie Boeynaems2, Bernard Robaye2, Catherine Bruyns2,
François Willermain1. 1Ophthalmology, Univ of Brussels-St Pierre Hosp, Brussels,
Belgium; 2Univ of Brussels-IRIBHM, Brussels, Belgium.
Purpose: To study the potential role of the nucleotide receptor P2Y2 in the
development of experimental autoimmune uveitis (EAU).
Methods: EAU were induced in WT and P2Y2 KO mice by direct immunization
with IRBP peptide 1-20 or adoptive transfer of in vitro restimulated semi-purified
lymphocytes from spleen and draining lymph nodes isolated from immunized mice.
Clinical and histological scores were used to grade disease severity. Lymphocyte
proliferation was measured by thymidine incorporation and cytokine secretion by
ELISA. Flow cytometry was used to analyze cell surface marker expression.
Results: EAU clinical scores were slightly decreased in KO mice. Similarly,
adoptive transfer of semi-purified lymphocytes from KO into C57Bl/6 WT mice
induced significantly less disease than the transfer of WT cells into C57Bl/6 WT
animals. Analysis of spleen and lymph node cells of immunized animals showed no
differences in term of cell populations. However, in vitro restimulation of KO cell
induced less proliferation and cytokine (IFN gamma, TNF alpha and IL-17)
secretion as compared to WT cells.
Conclusions: Our data show that P2Y2 mice are less susceptible to develop an
autoimmune response against IRBP peptide 1-20, influencing the development of
EAU. Those data are consistent with the hypothesis that P2Y2 play a role of danger
signal receptors.
Commercial Relationships: Laure E. Caspers, None; Lia J. Relvas,
None; Remi Dewispelaere, None; Maya Makhoul, None; Didier Communi,
None; Jean-Marie Boeynaems, None; Bernard Robaye, None; Catherine
Bruyns, None; François Willermain, None
Support: None
Program Number: 6237 Poster Board Number: D1097
Presentation Time: 8:30 AM - 10:15 AM
Role of iC3b-CR3 interaction in Experimental Autoimmune Anterior Uveitis
Bharati Matta, Purushottam Jha, Puran S. Bora, Nalini S. Bora. Ophthalmology,
Jones Eye Institute-UAMS, Little Rock, AR.
Purpose: To explore the role of interaction between complement activation product
- iC3b and its receptor CR3 in experimental autoimmune anterior uveitis (EAAU).
Methods: Male Lewis rats (5-6 weeks old) were immunized with bovine melanin
associated antigen (MAA) emulsified (1:1) in complete Freund’s adjuvant (CFA) in
the hind footpad. Animals were sacrificed on different days post-immunization and
lymphocytes were purified from the popliteal lymph nodes (LNs) using histopaque
gradient. Lymphocytes were stained with anti-CD4 APC and anti-iC3b followed by
staining with FITC labeled secondary antibody (Ab). In a separate set of
experiment, cells were stained with anti-CD11b/c PE, anti-CD3-APC, antiCD45RA-FITC and anti-iC3b followed by staining with PerCP labeled secondary
Ab. CFSE labeled lymphocytes purified from the popliteal LNs on day 5 postimmunization were plated in serum free CellGroR Medium containing rat iC3b in
the presence of different concentration of anti-CD11b and anti-iC3b Abs separately
for 5 days in the presence of bovine MAA (20 µg/ml). On day 6 non-adherent cells
were collected and stained with anti-CD4-APC. The proliferation data was
collected using FACS Calibur and was analyzed using Win MDI.
Results: During the induction of EAAU, iC3b is deposited on antigen presenting
cells (APCs) and CD4+ T cells present in the popliteal LNs. Out of total CD4+ T
cells, the percentage of CD4+ T cells that are positive for iC3b started to increase as
early as day 2, peaked on day 7 post-immunization and started to decrease after
that. Out of total CD11b/c+ CD3- CD45RA- cells, the percentage of cells that are
iC3b positive sharply increased at day 2 and peaked on day 7 post-immunization.
Treatment with anti-CD11b and anti-iC3b Abs separately inhibited the proliferation
of CD4+ T cells in response to MAA in a dose dependent manner.
Conclusions: Our results demonstrate that during the induction phase of EAAU,
iC3b is deposited on CD4+ T cells and CD11b/c+ CD3- CD45RA- APCs that are
present in the popliteal LNs of MAA sensitized Lewis rats. Our data further suggest
that the interaction between iC3b and its receptor CR3 modulates CD4+ T cells
responses in EAAU.
Commercial Relationships: Bharati Matta, None; Purushottam Jha,
None; Puran S. Bora, None; Nalini S. Bora, None
Support: NIH grant EY018812 and the grant from Pat and Willard Walker Eye
Research Center, Jones Eye Institute, University of Arkansas for Medical Sciences,
Little Rock, AR.
Program Number: 6238 Poster Board Number: D1098
Presentation Time: 8:30 AM - 10:15 AM
Temporal Expression of miR-155 Correlates with the initiation and
Development of Experimental Autoimmune Uveitis (EAU)
Bernadette Marrero, Yu Chen-Rong, Chandrasekharam Nagineni, Charles
Egwuagu. Immunology, NEI, Bethesda, MD.
Purpose: MicroRNAs are small non-coding genes that regulate diverse cellular
processes in vertebrates. Aberrant or enhanced expression of several miRNAs
including miR-146, miR-21, Let-7e and miR-155 are associated with several
pathogenic conditions of humans. For example, miR-155 has recently been
implicated in the pathogenesis of CNS inflammatory diseases such as multiple
sclerosis and experimental autoimmune encephalomyelitis (EAE). In this study, we
have investigated potential involvement of miR-155 in intraocular inflammatory
disease or uveitis.
Methods: To investigate the potential involvement of miR-155 in ocular
inflammatory diseases, we induced experimental autoimmune uveitis (EAU) in
C57BL/6 mice by active immunization with IRBP in CFA. Severity and
progression in EAU were assessed by fundoscopy and histopathology. miRNA
expression in human retinal cells (ARPE-19), as well as, in the retina of mice with
EAU was analyzed by real-time quantitative PCR.
Results: Analysis of inflammatory cells in the retina during EAU revealed a 5.3fold increase of miR-155 expression in the retina compared to un-immunized mice.
The increase in miR-155 expression correlated temporally with increase in proinflammatory cytokines and progression of EAU. In addition, culturing human RPE
cells in medium containing pro-inflammatory cytokines induced an even a more
dramatic increase in miR-155 compared to untreated cells.
Conclusions: Dramatic increase of miR-155 in the retina during ocular
inflammation and following exposure of ocular cells to inflammatory environment
was dramatic and suggest that miR-155 may be implicated in mechanisms that
promote of T cell-dependent tissue inflammation. Our data further suggest that
miR-155 might be a promising therapeutic target for the treatment of uveitis and
other autoinflammatory disorders.
Commercial Relationships: Bernadette Marrero, None; Yu Chen-Rong,
None; Chandrasekharam Nagineni, None; Charles Egwuagu, None
Support: None
Program Number: 6239 Poster Board Number: D1099
Presentation Time: 8:30 AM - 10:15 AM
DAP-12, a Major Immunomediator, Either Promotes or Suppresses EAU
Development
Barbara P. Vistica1, Vanessa Montalvo-Reddin1, Guangpu Shi1, Lindsey Nugent1,
Laura Quigley2, Daniel W. McVicar2, Igal Gery1. 1Lab of Immunology, National
Eye Institute, Bethesda, MD; 2Cancer and Inflammation Program, NCI-Frederick,
Frederick, MD.
Purpose: DAP12 (DNAX-activating protein of 12kDa), a transmembrane signaling
molecule, plays a major role in the immune response by transducing a variety of
activation signals from receptors on antigen-presenting and other leukocytes. The
role of DAP12 in the etiology of pathogenic autoimmunity has been controversial,
since DAP12 deficient mice reportedly exhibit either enhanced or depressed
experimental autoimmune diseases. Here, we compared DAP12 deficient mice and
wild type (WT) controls for their susceptibility to EAU induction and immune
responsiveness to the uveitogenic antigen
Methods: DAP12 deficient mice and their C57Bl/6 WT controls were obtained
from two animal facilities, on the same campus, both specific pathogen free (SPF)
but with different standards for helicobacter and other organisms. The mice were
immunized with interphotoreceptor retinoid-binding protein (IRBP) and their
development of EAU was evaluated by fundoscopy and histological examination.
Lymphoid cells from draining lymph nodes were cultured with IRBP to measure
proliferation and cytokine release.
Results: Although disease in control mice from both facilities was comparable
DAP12 deficient mice from the SPF facility with less stringent health standards
were superior to their WT controls in both the severity of their ocular inflammation
and the levels of proliferation and of IL-17 and interferon gamma produced by their
lymphocytes. In contrast, DAP12 deficient mice from the cleaner rodent barrier
facility were inferior to their controls by both parameters.
Conclusions: Our data provide a possible resolution to the controversy concerning
the role of DAP12 in the induction of EAU: deficiency in this molecule can either
promote or suppress the pathogenic response, even when the same experimental
system is used. The difference we observed between the two portions of the study
could be attributed only to conditions within the animal facilities that shape the
immune systems.
Copyright 2012 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2012 Annual Meeting Abstracts
Commercial Relationships: Barbara P. Vistica, None; Vanessa MontalvoReddin, None; Guangpu Shi, None; Lindsey Nugent, None; Laura Quigley,
None; Daniel W. McVicar, None; Igal Gery, None
Support: None
Program Number: 6240 Poster Board Number: D1100
Presentation Time: 8:30 AM - 10:15 AM
Inhibition of CdK5 Attenuates Experimental Autoimmune Uveitis
Zili Zhang1, Xiumei Wu1, Jie Duan1, James T. Rosenbaum2. 1Pediatrics, Oregon
Health & Science University, Portland, OR; 2Ophthalmology, Casey Eye InstituteOHSU, Portland, OR.
Purpose: T cells are essential for the development of autoimmune uveitis. Our
previous study showed that OX40, a key co-stimulatory molecule, plays a pivotal
role in enhancing T cell response and ocular inflammation. However, the molecular
mechanism by which OX40 regulates uveitogenic T cells remains to be fully
elucidated. Recently, cyclin dependent kinase 5 (CdK5), a unique serine/threonine
kinase, has been implicated in T cell activation. Furthermore, CdK5 exerts several
key downstream effects in common with OX40. Thus, we sought to determine
whether CdK5 is a critical intermediary of OX40 signaling, leading to exacerbation
of uveitis.
Methods: The splenocytes of DO11.10 mice were activated by cognate antigen
ovalbumin (OVA) for up to 72 hours. Flow cytometry was performed to compare
intracellular CdK5 expression between CD4+OX40+ and OX40- T cells. In
addition, these cells were treated with CdK5 inhibitor Roscovitine, and T cell
apoptosis was measured by Annexin V staining and MitoCapture assay. Lastly,
experimental autoimmune uveitis (EAU) was induced by immunization of B10.RIII
mice with interphotoreceptor binding protein. Some mice were further treated with
OX40 activating antibody, and EAU score was determined on day 21 to assess the
effect of Roscovitine on OX40-enhanced uveitis.
Results: To explore the relationship of OX40 with CdK5, we categorized DO11.10
CD4+ cells to 2 distinct populations based on surface OX40 expression. After OVA
activation, OX40+ cells expressed more intracellular CdK5 than OX40lymphocytes. In addition, blocking phosphatidylinositol 3-kinases, which mediates
OX40 signaling pathway, reduced the expression of CdK5. Moreover, inhibition of
CdK5 by Roscovitine led to the decrease of activated CD4+CD44+ lymphocyte
number, which coincided with increased T cell apoptosis. Finally, weekly treatment
of Roscovitine attenuated the severity of OX40-enhanced EAU.
Conclusions: These results implicated CdK5 in OX40-augmented T cell response
and suggest that targeting CdK5 may be a novel therapeutic strategy for treating T
cell-mediated uveitis.
Commercial Relationships: Zili Zhang, None; Xiumei Wu, None; Jie Duan,
None; James T. Rosenbaum, None
Support: NIH grant EY016788
Program Number: 6241 Poster Board Number: D1101
Presentation Time: 8:30 AM - 10:15 AM
Immunological Inhibition of Pigment Epithelium-Derived Factor (PEDF) ?
Charles E. Thirkill. Ocular Immunology Research Lab 1220 Surge III, UC Davis,
Davis 95616, CA.
Purpose: The biological activity of Pigment Epithelium-Derived Factor (PEDF,
molecular weight 45 kd) includes the inhibition of the activity of Vascular
Endothelial Growth Factor (VEGF,molecular weight 45 kd with smaller isoforms).
Both of these ubiquitous factors are multi-functional. Of relevance to this study is
the recognized expression of PEDF and VEGF within the retinal pigment
epithelium. The combination of these two factors normally contributes to the
balance of activity that maintains ocular homeostasis. Any immunological
inhibition of PEDF might result in an imbalance encouraging the blood vessel
propagation typical of several diseases that involve uncontrolled vascular
proliferation, such as that seen in some cancers, diabetic retinopathies and AgeRelated Macular Degenerations. A search for evidence that might implicate the
immunological inhibition of PEDF was performed on cancer patients presenting
with evidence of the 45 kd CAR syndrome. Rationale: PEDF shares the same
molecular mass as the unidentified 45 kd CAR antigen raising suspicion they might
be one and the same molecule. The possibility of immunological inhibition of the
activity of PDEF might then be of significance in both tumor growth, and its role in
maintaining introcular equilibrium. The option that the 45 kd CAR antigen may be
the 45 kd glycoprotein VEGF was included in these inquiries, but the potent
antibody activity against this antigen did not support the likelihood.
Methods: Western blot analyses were performed on the patient’s antibody activity
with the protein components of pig retina, and in vitro cultivated retinal pigment
epithelium (ATCC ARPE-19 CRL-2302). The 45 kd component of retinal pigment
epithelium was immunoprecipitated from an extract of cultured RPE using 45 kd
reactive cancer patient’s serum, and the precipitate subjected to proteomic analysis.
Results: Preliminary proteomic analyses implicate pigment epithelium-derived
factor as the 45 kd antigen of interest.
Conclusions: The possibility of an antibody-mediated interferece in the biolocal
activity of pigment epithelium-derived factor leads to a testable hypothesis:
“Immunological inhibition of the modulating anti-vascular proliferation properties
of pigment epithelium-derived factor results in a loss of homeostasis leading to the
excessive production of blood vessels that typifies some forms of cancer, and
retinopathies that involve uncontrolled vascular propagation”. If this hypothesis
proves correct it will introduce the prospect of ameliorating the pathological
process of unwanted vascular spread through approriate targeted
immunomodulation therapy.
Commercial Relationships: Charles E. Thirkill, Athena Diagnostics (P)
Support: RPB and NEI core grant 1P30 EY12576-7
Program Number: 6242 Poster Board Number: D1102
Presentation Time: 8:30 AM - 10:15 AM
Label-free LC-MSMS-based Differential Proteome Analysis of Vitreous from
Autoimmune Uveitis Cases
Stefanie M. Hauck1, Florian Hofmaier2, Johannes Dietter1,3, Marcel Blindert1,
Elisabeth Kremmer4, Margarete E. Swadzba2, Barbara Amann2, Cornelia A. Deeg2,
Marius Ueffing1,3. 1Department of Protein Science, Helmholtz Center Munich,
Neuherberg, Germany; 2Department for Veterinary Sciences, Institute of Animal
Physiology, Munich, Germany; 3Centre for Ophthalmology, Institute for
Ophthalmic Research, Tubingen, Germany; 4Institute for Molecular Immunology,
Helmholtz Center Munich, Munich, Germany.
Purpose: Equine recurrent uveitis (ERU) is a devastating immune mediated
inflammation targeting the inner eye and ultimately leading to blindness in the
affected animals. The widespread disease (10%) is characterized by recurring
episodes of inflammation followed by quiescent stages. ERU represents the only
spontaneous animal model for human autoimmune uveitis and thus serves as a
valuable model for exploration of as yet unknown pathomechanisms relevant for
disease progression and relapses. Since vitreous is directly adjacent to the inflamed
retina this compartment facilitates indirect exploration of ongoing disease related
events in the retina. To investigate protein expression changes related to the
disease, we undertook a comprehensive differential proteome profiling of ERU and
healthy vitreous.
Methods: Proteins from vitreous samples of healthy horses and ERU cases (n=16)
were subjected to tryptic digestion and analysed by liquid chromatography mass
spectrometry (LC-MSMS; OrbiTrap). All detected peptide features were aligned
and statistically analysed for differential abundance based on cumulated peak
intensities. Peptides were identified by database search (Ensembl, Equus Caballus)
and resulting protein IDs were grouped according to their mode of regulation.
Pathway enrichment analyses (Consensus PathDB and Genomatix Pathway
Systems) were used for identification of overrepresented pathways per condition.
Candidate proteins were validated by western blots on a large sample collection.
Results: Highly sensitive comparisons of vitreous samples resulted in identification
of 251 proteins, 105 of these were identified with very high confidence (2 or more
peptides, p<0.01). 26 proteins were found upregulated in ERU cases, while 44 were
downregulated. Pathway enrichment analyses with these regulated proteins resulted
in the identification of several significantly overrepresented pathways, among them
“ECM-receptor interaction” (KEGG) and “Wnt signaling” (INOH). From these
pathways, selected candidates were validated by western blots and mode of
regulation observed by quantitative mass spectrometry was confirmed in all cases.
Interaction network analyses based in literature mining enabled to suggest potential
pathomechanisms and will be discussed in detail.
Conclusions: Quantitative label-free proteomics based on LC-MSMS identified a
signature of protein markers in the vitreous of uveitic eyes that point to dynamic
changes in both extracellular matrix homeostasis and wnt signalling. As both
molecular networks are mechanistically linked to inflammation as well as vascular
changes in the eye, members of these protein networks are likely involved in the
pathogenesis of uveitis.
Commercial Relationships: Stefanie M. Hauck, None; Florian Hofmaier,
None; Johannes Dietter, None; Marcel Blindert, None; Elisabeth Kremmer,
None; Margarete E. Swadzba, None; Barbara Amann, None; Cornelia A. Deeg,
None; Marius Ueffing, None
Support: German Federal Ministry of Education and Research: HOPE - FKZ
01GM0852; SFB 571 A5 Deeg and DFG De 719/2-2.
Program Number: 6243 Poster Board Number: D1103
Presentation Time: 8:30 AM - 10:15 AM
Hsa-mir-let 7i Treatment Suppresses Tlr4 Mediated Mitochondrial Oxidative
Stress In Giant Cell Arteritis
Narsing A. Rao, Sindhu Saraswathy. Ophthalmology, Doheny Eye Institute, Los
Angeles, CA.
Purpose: Our previous findings have shown downregulation of 15 miRNAs related
to TLR4, its signaling molecules, and mitochondrial oxidative stress in giant cell
arteritis (GCA). One of the 15 miRNAs, hsa-let-7i is validated to target TLR4. In
this study, we investigate the role of hsa-let-7i in the modulation of TLR4 and
mitochondrial oxidative stress in GCA that lead to amplification of the
inflammatory process.
Methods: Non-GCA temporal artery biopsy specimens were exposed to LPS and
Copyright 2012 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2012 Annual Meeting Abstracts
subjected to total RNA isolation and immunohistochemical studies to detect TLR4,
CD209 (dendritic cells) and CD31 (endothelial cells). qPCR analysis was
performed to detect TLR4 and CD83 gene expression. miRNA PCR Array
(SABiosciences, Frederick, MD) was performed using specific primers for hsa-mirlet 7i, mir-let 7a, mir-10a, mir-106b, mir-221. Another group of similar specimens
were treated with miRNA hsa-let-7i mimic and LPS. Scrambled miRNA and
samples without LPS treatment were used as controls. TLR4 expression was
studied by qPCR analysis. Another set of specimens were exposed to LPS and then
treated with the TLR4 inhibitor OXPAPC or miRNA hsa-let-7i mimic or scrambled
microRNA. DNA damage marker 8-OHdG was colocalized with the mitochondrial
marker COX IV by immunohistochemistry.
Results: TLR4 was localized in the dendritic cells in the adventitia region of the
temporal artery after LPS exposure whereas it was absent on the endothelial layer.
miRNA-hsa-let 7i in LPS treated temporal artery specimens were significantly
downregulated compared to non-treated controls.TLR4 was markedly upregulated
in the temporal artery exposed to LPS. Such overexpression was suppressed with
miRNA hsa-let-7i mimic treatment whereas it was not altered with scrambled
microRNA treatment. 8-OHdG was colocalized with COX-IV in the adventitia of
the temporal artery exposed to LPS. Such damage was absent in the artery treated
with the TLR4 inhibitor or with miRNA hsa-let-7i mimic.
Conclusions: The increased TLR4 expression on the dendritic cells of adventitia
indicates that the initial inflammatory insult in GCA could begin in the adventitia.
This study reveals that TLR4 activation from LPS leads to mitochondrial oxidative
stress, and such TLR4 mediated damage can be prevented by the miRNA hsa-let-7i
mimic treatment.
Commercial Relationships: Narsing A. Rao, None; Sindhu Saraswathy, None
Support: NIH grants: EY017347, EY019506, EY03040 and RPB
Program Number: 6244 Poster Board Number: D1104
Presentation Time: 8:30 AM - 10:15 AM
Amelioration of Experimental Autoimmune Uveoretinitis by Inhibiton of
Toxic AGEs Formation
Zhenyu Dong1A,1B, Nobuyoshi Kitaichi2,1B, Daiju Iwata1A,1B, Ryo Ando1A,1B, Junichi
Fukuhara1A,1B, Anton M. Lennikov1A,1B, Atsuhiro Kanda1A,1B, Kousuke Noda1A,1B,
Shigeaki Ohno1C, Susumu Ishida1A,1B. ADepartment of Ophthalmology, BLaboratory
of Ocular Cell Biology and Visual Science, CDepartment of Ocular Inflammation
and Immunology, 1Hokkaido University Graduate School of Medicine, Sapporo,
Japan; 2Department of Ophthalmology, Health Sciences University of Hokkaido,
Sapporo, Japan.
Purpose: Advanced glycation end products (AGEs) are permanently modified
protein derivatives, forming through non-enzymatic glycation of amino groups.
Since glyceraldehyde-derived AGEs have strong toxicity, it is considered as toxic
AGEs (TAGE). TAGE has been demonstrated to play an important role in the
pathogenesis of chronic inflammatory diseases through interaction with its receptor
for AGEs (RAGE). Pyridoxamine, one of the vitamin B6 derivatives, has been
demonstrated to inhibit the Amadori rearrangement in the process of AGEs
production. In this study, we quantified the serum TAGE levels in patients with
endogenous uveitis. In addition, therapeutic effects of pyridoxamine were
examined on experimental autoimmune uveoretinitis (EAU), a murine model of
human endogenous uveitis.
Methods: One hundred three consecutive patients of uveitis (31 VKH disease
patients in convalescent stage, 21 with HLA-B27 associated uveitis, 14 with Behcet
disease, and 37 with sarcoidosis) and 33 healthy volunteers were enrolled. Serum
TAGE levels of participants were quantified with ELISA. In EAU experiments,
female B10.BR (H-2k) mice were immunized with interphotoreceptor retinoidbinding protein (IRBP)-derived peptide emulsified with complete Freund’s
adjuvant containing M.tuberculosis, and continuously administered 200mg/kg or
400mg/kg of pyridoxamine every 24 hours orally. Severity of EAU was evaluated
both clinically and histopathologically. Translocation of NF-κB p65 into nucleus,
and retinal levels of TAGE were also examined immunohistochemically.
Results: Serum TAGE levels of uveitis patients with each etiology were
significantly higher than those of healthy controls (P<0.001 for all). EAU was
significantly milder both clinically and histopathologically when treated with
400mg/kg of pyridoxamine (P<0.05). TAGE levels were decreased in both sera and
retina (P<0.05 for both), and translocation of NF-κB p65 into nucleus was downregulated in retinas in pyridoxamine-treated EAU mice.
Conclusions: TAGE levels were upregulated in sera of uveitis patients, and
systemic administration of pyridoxamine ameliorated EAU. Our results suggest
that TAGE-RAGE system contributes to the phathogenesis of uvetitis and is a
novel therapeutic target for endogenous uveitis.
Commercial Relationships: Zhenyu Dong, None; Nobuyoshi Kitaichi,
None; Daiju Iwata, None; Ryo Ando, None; Junichi Fukuhara, None; Anton M.
Lennikov, None; Atsuhiro Kanda, None; Kousuke Noda, None; Shigeaki Ohno,
None; Susumu Ishida, None
Support: None
Program Number: 6245 Poster Board Number: D1105
Presentation Time: 8:30 AM - 10:15 AM
Ophthalmological and Histological Manifestations of IgG4-Related Disease
Elise Philippakis, Valerie Touitou, Coralie Bloch-Queyrat, Aude Rigolet, Frederic
Charlotte, Phuc LeHoang, Bahram Bodaghi. Pitie Salpetriere Hospital, Paris,
France.
Purpose: IgG4-related disease is defined by both elevated serum IgG4 over
135mg/dL and histopathological features, such as lymphocytes and IgG4+ plasma
cell infiltration (ratio IgG4+ plasma cells/IgG+ plasma cells over 40%), associated
with typical tissue fibrosis or sclerosis. The aim of this study is to determine the
ophthalmological and histocytological features of IgG4-related disease.
Methods: Patients with biopsy-proven hyper-IgG4 syndrome, seen between 2009
and 2011 in a single tertiary center, were retrospectively reviewed. Clinical features
were analyzed, and compared with histological features and with patient's
prognosis.
Results: Three patients were included in the study. Mean age at diagnosis was 58
years (range:50-63 years). One patient presented with unilateral painful propotosis,
associated with third and fifth cranial nerves palsies. One patient had recurrent
anterior scleritis, and one patient presented with chronic conjonctival infiltration.
None of them had extra-ocular involvement (FDG-PET-scan). Other inflammatory
conditions were ruled out. Histocytology demonstrated lymphoplasmocytosic
proliferation with overexpression of IgG4+ plasma cells and signs of fibrosis. All
patients responded to corticosteroids. However one patient developped orbital
fibrosis and required the adjunction of an immusuppressive drug.
Conclusions: IgG4 related disease is a new and rare entity, which can be
challenging to diagnose when clinical features are atypical , such as isolated
conjonctivitis, scleritis and orbital infiltrations. Histopathology is the gold-standard
for the diagnosis. Prognosis is determined by fibrosis, sclerosis and systemic
involvement(pancreatitis, retroperitoneal fibrosis and hepatitis).Treatment is not
consensual, but is based on corticosteroids, associated most of the time with
immunosuppressants in case of fibrosis or cortico- dependant lesions.
Commercial Relationships: Elise Philippakis, None; Valerie Touitou,
None; Coralie Bloch-Queyrat, None; Aude Rigolet, None; Frederic Charlotte,
None; Phuc LeHoang, None; Bahram Bodaghi, None
Support: None
Program Number: 6246 Poster Board Number: D1106
Presentation Time: 8:30 AM - 10:15 AM
Ocular Immune Pathological Analysis in a Murine Model of Anterior Scleritis
Hiroko Taniguchi1, MingCong Wang1, Atsuo Nakajima2, Junko Hori1.
1
Ophthalmology, Nippon Medical School, Tokyo, Japan; 2Rheumatology, Tokyo
Metropolitan Police Hospital, Tokyo, Japan.
Purpose: As the sclera comprises type II collagen, scleritis is often associated with
autoimmune diseases such as rheumatoid arthritis. We have previously reported the
establishment of an anterior scleritis model through modification of a collageninduced arthritis model. The present study conducted ocular immunological
analysis in this same model to investigate the scleritis pathology.
Methods: Arthritis and anterior scleritis were induced in 8-week-old DBA/1J mice
via subcutaneous injection of bovine type II collagen emulsified with complete
Freund’s adjuvant into the posterior neck (primary immunization) and 21 days later
into the periocular region (secondary immunization). Eyeballs were excised at 3, 5
and 8 weeks after secondary immunization and analyzed histologically and
immunohistologically.
Results: Clinical findings comprised severe arthritis and dilation of scleral blood
vessels. Anterior scleral thickening was observed histologically, with significantly
more inflammatory infiltrating cells in the anterior sclera compared to control mice.
Infiltration of CD4+ and CD11b+ cells was observed, but no CD11c+ cells were
present, while deposition of plasma cells (CD138), complement (C3) and
immunoglobulin (IgG and IgM) was particularly evident in the region of the
anterior sclera in contact with the ciliary body.
Conclusions: T cells, macrophages, plasma cells, complement and
immunoglobulins were present in the sclera of the collagen-induced anterior
scleritis model, suggesting the involvement of immunocomplex deposition in the
induced scleritis of the present model.
Commercial Relationships: Hiroko Taniguchi, None; MingCong Wang,
None; Atsuo Nakajima, None; Junko Hori, None
Support: Grants-in-Aid for Scientific Research (C) from Japan Society for the
Promotion of Science
Program Number: 6247 Poster Board Number: D1107
Presentation Time: 8:30 AM - 10:15 AM
Erythrocyte Sedimentation Rate and C-Reactive Protein in Anterior Uveitis
Justin D. Marsh, Bethany B. Markowitz. University of South Carolina, Columbia,
SC.
Purpose: Erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) are
laboratory tests used for monitoring disease processes due to their utility as nonspecific markers of inflammation. Not infrequently, systemic inflammatory
processes show evidence of anterior segment inflammation. This study aimed to
Copyright 2012 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2012 Annual Meeting Abstracts
evaluate ESR and CRP values during periods of anterior uveitis to determine if
ocular inflammation was sufficient to elevate these laboratory values to levels
considered outside the normal range.
Methods: A retrospective study was performed by reviewing patient charts with
ICD-9 diagnosis codes of anterior uveitis (364.01, 364.02, 364.04, 364.3) seen at
University of South Carolina Ophthalmology Specialty Clinics over a two year
period. From within this subset, charts were further reviewed to select patients in
which there was a recorded ESR or CRP value within seven days of documented
anterior uveitis by an attending ophthalmologist or resident within the department.
Sixteen patients were found that met these criteria, all of which had ESR values
within one week of examination. Five of the sixteen patients also had CRP values
documented within one week of examination. Additional data including age, sex,
degree of inflammation, etiology of inflammation, and bilaterality were recorded as
documented. In cases of bilateral anterior uveitis, the degree of inflammation was
determined by the more severe eye. ESR and CRP values were then stratified based
on reference range values. ESR was considered within the normal range when < 20
mm/hr for patients of age < 40. For patients of age > 40, the normal reference range
was calculated as < Age/2 for males, and < (Age +10)/2 for females. CRP was
considered within the normal reference range when < 10 mg/L regardless of age or
sex.
Results: Sixteen patients were evaluated, two of which had multiple examinations
corresponding with laboratory data. ESR and CRP were found to be within the
normal reference range in 57% (12/21) and 60% (3/5) of patients, respectively. In
patients with laboratory data on the same day as examination, ESR and CRP were
within the normal reference range in 55% (6/11) and 67% (2/3), respectively. In
patients with at least 2+ cellular reaction documented, ESR was found to be within
the normal reference range in 60% (6/10) of patients.
Conclusions: While ESR and CRP may be elevated in multiple inflammatory
processes, anterior uveitis alone does not appear to be sufficient to yield elevated
values beyond the normal reference range. In this study, ESR and CRP were within
the normal range in 57% and 60% of patients with anterior uveitis, respectively.
Commercial Relationships: Justin D. Marsh, None; Bethany B. Markowitz,
None
Support: None
Program Number: 6248 Poster Board Number: D1108
Presentation Time: 8:30 AM - 10:15 AM
Antiphospholipid syndrome in Mexican population
Claudia Recillas-Gispert, Juan Carlos Martinez. Ophthalmology, INCMNSZ,
Mexico City, Mexico.
Purpose: To determine the prevalence of Antiphospholipid syndrome in the
Mexican population over 10 years, and the frequency of ocular manifestations, and
which of those cases were the first manifestation of the disease.
Methods: Retrospective, observational, transversal and descriptive study reviewing
the medical files and records from all patients with Antiphospholipid syndrome’s
diagnostic with descriptive statistical analysis for age, gender, type of
antiphospholipid syndrome (primary or secondary), ocular manifestations and
visual acuity at our hospital in a period of 10 years.
Results: We include 332 patients with diagnostic of Antiphospholipid syndrome.
Mean age at the moment of the diagnostic was 29.54 years. 278 were women
(83.73%). 141 patients (42.85%) were diagnosed with Primary Antiphospholipid
syndrome (PAPS), which 9 patients (6.38%) were ocular PAPS; 191 patients
(57.17%) with Secondary Antiphospholipid syndrome (SAPS), 14 patients (7.33%)
presented one ocular thrombotic episode without other systemic episode. Patients
with ocular vasso-oclusive manifestations were 15 with central vein vascular
oclussion (CRVO):8, central arteriy vascular oclusion (CAVO): 6, branch retinal
artery oclusion: 1, optic neuritis:12, retinal vasculitis:7 amaurosis fugax:3. Other
ocular autoimmune manifestations: epiescleritis: 2, escleritis:2, papiledema:1,
internuclear ophthalmoplegic: 1, uveitis:1.Visual prognosis in patients with ocular
manifestations presented a wide range associated to the type of alteration. Patients
with legal blindness in PAPS were 3: 2 related to CRAO and the other one with
optical neuritis, while patients with SAPS were 6: 2 with CRVO, 2 retinal
vasculitis, 1 with CRAO and 1 with optic neuritis.
Conclusions: Ocular manifestations in antiphospholipid syndrome have a low
prevalence with a tendency to be more affected in the SAPS, may be because of
comorbility with other diseases. However, they have great impact on the visual
prognosis, so it is important to do timely assessment on suspected APS patients.
Commercial Relationships: Claudia Recillas-Gispert, None; Juan Carlos
Martinez, None
Support: None
Eye Research and Surgery Institution, Cambridge, MA; 3Ophthalmology,
BayWiew Clinic, Mumbai, India; 4Ophthalmology, Tauber Eye Center, Kansas
City, MO; 5Ophthalmology, Harvard Medical School, Boston, MA.
Purpose: To evaluate the demographic characteristics, clinical features, ocular
complications, and successful therapeutic regimens in patients with scleritis
associated with inflammatory bowel disease (IBD).
Methods: Retrospective case series. We reviewed the electronic health records of
500 patients with scleritis seen at two tertiary referral centers and selected the ones
with IBD. Patient characteristics, clinical features, ocular complications and
successful therapeutic regimens were evaluated.
Results: Of 500 patients with scleritis, 11 patients had IBD (2.2%), 9 with Crohn´s
disease (CD) and 2 with ulcerative colitis (UC); that includes 8 women and 3 men
with a mean age of 47 years (range, 33 to 67 years). Six of the patients with CD
had diffuse scleritis, 1 had nodular scleritis, 1 had necrotizing scleritis, and 1 had
necrotizing scleritis and posterior scleritis; anterior uveitis was present in 2
patients, peripheral keratitis in 3 patients, and glaucoma in 1 patient but final visual
acuity was not decreased in any patient. Scleritis occurred after several episodes of
anterior uveitis in 2 patients. The 2 patients with UC had diffuse scleritis with
anterior uveitis without corneal lesions or glaucoma or decrease of vision. There
was no previous uveitis. Scleritis was the initial manifestation whose study led to
the diagnosis of CD in 4 of the 9 patients and to the diagnosis of UC in all 2
patients. All 11 patients had arthritis and had recurrent scleritis. Successful
therapeutic regimens in scleritis with CD included 1 patient with sulfasalazine, 1
patient with intravenous steroids, 3 patients with methotrexate, 2 patients with
azathioprine, and 2 patients with infliximab. Successful therapeutic regimens in
scleritis with UC included 1 patient with infliximab and 1 patient with sirolimus.
Conclusions: Scleritis associated with IBD is more common in CD than in UC. It
is usually recurrent and may take the form of any type of scleritis, including
necrotizing and posterior scleritis. Although ocular complications may appear, they
unfrequently cause decrease of vision. Patients who develop scleritis after recurrent
anterior uveitis should be examined for CD. Scleritis associated with IBD is more
common in patients with arthritis and may be the initial manifestation of CD or UC.
Scleritis associated with IBD most often will require immunomodulatory therapy or
biologic response modifier drugs.
Commercial Relationships: Maite Sainz de la Maza, None; Nicolas Molina,
None; Luis A. Gonzalez-Gonzalez, None; Priyanka P. Doctor, None; Joseph
Tauber, None; Stephen Foster, None
Support: None
Program Number: 6250 Poster Board Number: D1110
Presentation Time: 8:30 AM - 10:15 AM
Posterior Scleritis and Orbital Mass Associated to Positive Antineutrophil
Cytoplasmic Autoantibodies Without Systemic Involvement
MarÃa de los Angele Ramos Cadena1, Gustavo Aguilar Montes1, Matilde Ruiz
Cruz2. 1Ophthalmology, Hospital General Dr. Manuel Gea Gonzalez, Mexico City,
Mexico; 2Ophthalmology, Centro de Investigacion de Enfermedades Infecciosas
del Instituto Nacional de Enfermedades Respiratorias, Mexico City, Mexico.
Purpose: To present a clinical case of a 16 year-old Mexican patient, without any
relevant previous medical history and systemic symptoms, diagnosed with posterior
scleritis associated to positive antineutrophil cytoplasmic autoantibodies (ANCA)
directed to Proteinase 3 (PR3), with clinical involvement of the right eye.
Methods: We performed a complete ophthalmologic examination, laboratory and
immunologic tests, ocular ultrasound, orbit and paranasal sinus CT Scan and
cranial MRI.
Results: Funduscopic examination revealed papillitis, macular scar, serous retinal
detachment, and vitritis. Ocular ultrasound of the right eye showed T-sign,
enlargement of the posterior wall and intraorbital optic nerve. Laboratory results
did not show renal or other systemic involvement. ANCA results showed positive
cANCA, and Anti-PR3. CT Scan revealed a small mass in the orbit cone, close to
the optic nerve entrance, and enlargement of the intraorbital portion of the optic
nerve. MRI disclosed a normal intracraneal optic nerve.
She was treated with intravenous methylprednisolone 1g per day, during three days,
followed by oral prednisone at immunosuppressive dose, tapered according to
evolution, discontinued after one month and a half. The ophthalmologic and
imagenologic manifestations ceased after the first month of treatment.
Conclusions: 50% of posterior scleritis are related to systemic involvement. This
patient presented positive cANCA and Anti-PR3 even after corticosteroid
treatment, which would usually imply systemic activity; but in this case the disease
presented without extraocular manifestations. Immunosuppressive treatment should
be indicated if systemic involvement occurs or ocular symptoms recur because of
Program Number: 6249 Poster Board Number: D1109
Presentation Time: 8:30 AM - 10:15 AM
Scleritis Associated with Inflammatory Bowel Disease
Maite Sainz de la Maza1, Nicolas Molina1, Luis A. Gonzalez-Gonzalez2, Priyanka
P. Doctor3, Joseph Tauber4, Stephen Foster2,5. 1Instituto Clinico Oftalmologia,
Hospital Clinico Oftalmologia, Barcelona, Spain; 2Ophthalmology, Massachusetts
Copyright 2012 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2012 Annual Meeting Abstracts
the high risk of vasculitic multiorgan affection and renal failure.
Commercial Relationships: MarÃa de los Angele Ramos Cadena,
None; Gustavo Aguilar Montes, None; Matilde Ruiz Cruz, None
Support: None
Program Number: 6251 Poster Board Number: D1111
Presentation Time: 8:30 AM - 10:15 AM
Uveitis In Patients With Diabetes Mellitus
Martha C. Fuentes, Cintia G. Sánchez-Balleza, Miguel Pedroza-Seres. Ocular
Inflammation and immunology, CONVAL, Mexico, Mexico.
Purpose: 1) Describe diabetes frequency in population with uveitis attending
Conde de la Valenciana Institute, 2) describe demographic characteristics and 3)
asses the relationship between hyperglycemia and inflammatory reaction in patients
with diabetes and active uveitis.
Methods: Prospective case series, observational and descriptive. We recruited
patients with active uveitis and diabetes mellitus. The initial evaluation of patients
was conducted by two researchers and included complete ophthalmic history, best
corrected visual acuity (BCVA), slit lamp biomicroscopy and fundus evaluation.
The degree of inflammatory activity was determined according to SUN. Central
glucose was requested at the time of diagnosis of uveitis or reactivation and during
every visit. Comparison of blood glucose and degree of inflammatory activity was
performed for all the observation points. Glycemia was related to the degree of
inflammatory activity using Spearman test.
Results: 315 patients with uveitis were evaluated, 48 patients were diabetic
(15.2%), two patients had type 1 diabetes (2%). We found 38 female (79.1%),
unilateral uveitis presented in 27 cases (56.2%); mean age was 56.75, time course
of diabetes was 10.64 years. The most frequent cuases of uveitis were nongranulomatous anterior uveitis 29.1% (n=14), panuveitis 12.5% (n=6), VKH
12.5% (n=6), herpetic uveitis 10.4% (n=5) and scleritis 10.4% (n=5). Twenty five
(51.02%) patients presented its first episode of uveitis. There is a statistically
significant positive correlation (p<0.05) between the anterior chamber
inflammation and hyperglycemia in non granulomatous idiopatic anterior uveitis
and herpetic uveitis.
Conclusions: Non granulomatous anterior uveitis, VKH and panuveitis are the
most frequent types of uveitis in patients with diabetes. There is a positive
correlation between hyperglycemia and inflammation in anterior chamber in
patients with anterior uveitis. The behavior of uveitis in these patients is more
aggressive and occurs more often bilaterally.
Commercial Relationships: Martha C. Fuentes, None; Cintia G. SánchezBalleza, None; Miguel Pedroza-Seres, None
Support: None
Program Number: 6252 Poster Board Number: D1112
Presentation Time: 8:30 AM - 10:15 AM
Risk Factors Associated with the Relapse of Uveitis in Patients with Juvenile
Idiopathic Arthritis
Ujwala H. Baheti1, Alaa Radwan1, Cheryl Arcinue1, Ravi Parikh1, Ashik
Mohamed2, C Stephen Foster1. 1Ophthalmology, Massachusetts Eye Research and
Surgery Institution, Cambridge, MA; 2Hyderabad Eye Research Foundation, L V
Prasad Eye Institute, Hyderabad, India.
Purpose: To identify risk factors associated with relapse of uveitis in patients with
juvenile idiopathic arthritis (JIA) - associated chronic or recurrent uveitis following
treatment with immunomodulatory therapy (IMT) and a drug free remission of 1
year.
Methods: Retrospective chart review of 30 patients with JIA associated uveitis,
who were successfully treated with IMT to a state of corticosteroid-free remission,
and subsequently remained in remission following discontinuation of IMT for a
period of at least 1 year. In subsequent follow up, some patients had relapse of
uveitis while others continued to be in remission. Remission was defined as <1+
cells in the anterior chamber or vitreous. Relapse was defined as > 1+ cells in the
anterior chamber or vitreous. We compared patients in remission with those who
relapsed, in an effort to evaluate risk factors associated with the relapse.
Results: Out of 30 patients, 17 (56.7%) remained in remission while 13 (43.3%)
relapsed. The patients in the remission group received IMT earlier (median of 9
months, inter-quartile range [IQR] 6-12 months) in the course of disease from
diagnosis as compared to patients in the relapse group (median of 78 months, IQR
36-120 months) (Mann-Whitney test, p = 0.0004). Moreover, the patients in the
remission group had received treatment with IMT at a younger age (median age 6
years, IQR 4-9 years) as compared to the relapse group (median age 11years, IQR
8-15 years) (Mann-Whitney test, p = 0.04). None of the other factors studied
revealed significant association with the relapse of uveitis.
Conclusions: Patients with uveitis associated with JIA treated with IMT earlier in
the course of disease and at a younger age have less likelihood of relapse of the
uveitis following discontinuation of the IMT after a 2 year period of quiescence on
IMT.
Commercial Relationships: Ujwala H. Baheti, None; Alaa Radwan,
None; Cheryl Arcinue, None; Ravi Parikh, None; Ashik Mohamed, None; C
Stephen Foster, None
Support: None
Program Number: 6253 Poster Board Number: D1113
Presentation Time: 8:30 AM - 10:15 AM
Risk Factors for Loss of Visual Acuity among Patients with Uveitis Associated
With Juvenile Idiopathic Arthritis: The SITE Study
Jennifer E. Thorne1, Anthony Gregory1, Ebenezer Daniel2, C Stephen Foster3,
Douglas A. Jabs4, Grace A. Levy-Clarke5, Robert B. Nussenblatt6, James T.
Rosenbaum7A, Eric B. Suhler7B, John H. Kempen8. 1Ophthalmology, Johns Hopkins
Wilmer Eye Inst, Baltimore, MD; 2Ophthalmology, University of Pennsylvania,
Philadelphia, PA; 3Ophthalmology, Ocular Immunol & Uveitis Fndtn, Cambridge,
MA; 4Ophthalmology, Mount Sinai School of Medicine, New York, NY; 5Tampa
Uveitis, Tampa, FL; 6National Eye Inst/NIH, Bethesda, MD; AOphthalmology,
B
Uveitis Clinic/Portland VAMC, 7Casey Eye Institute-OHSU, Portland, OR;
8
Ophthal-Biostatistics & Epidemiol, Scheie Eye Inst/Univ of Penn, Philadelphia,
PA.
Purpose: To describe the incidence of and risk factors for ocular complications and
loss of visual acuity (VA) in patients with juvenile idiopathic arthritis (JIA)associated uveitis.
Methods: 327 patients (599 affected eyes) with JIA-associated uveitis managed at
five tertiary ocular inflammation clinics in the United States were identified from
the Systemic Immunosuppressive Therapy for Eye Diseases (SITE) Cohort Study.
Demographic and clinical characteristics were obtained for every eye of every
patient at every visit via medical record review by trained expert reviewers. Loss of
VA to 20/50 or worse and to 20/200 or worse and the development of ocular
complications were assessed.
Results: At presentation, 240 (40.3%) eyes had a VA of 20/50 or worse; 144
(24.2%) had a VA of 20/200 or worse; and 359 (60.2%) had at least one ocular
complication. Over a median follow up of 3 years (range = 1 month to 25 years),
the rates of VA loss to the 20/50 or worse and 20/200 or worse thresholds were
0.18 and 0.09 per eye-year (EY), respectively. Among the affected eyes, the
incidence of developing at least one new ocular complication over follow up was
0.15/EY (95% confidence interval [CI]: 0.13/EY, 0.17/EY). However, among eyes
with uveitis that had no complications at presentation, the rate of developing at
least one ocular complication during follow up was 0.07/EY (95% CI: 0.04, 0.11).
After controlling for confounding factors, presence of posterior synechiae, active
uveitis, and prior intraocular surgery were statistically significantly associated with
VA to the 20/50 or worse and 20/200 or worse thresholds at presentation and
during follow up. Increasing anterior chamber cell was associated with increased
rates of visual loss in a dose-dependent fashion. Use of immunosuppressive drugs
reduced the risk of visual loss, particularly for the 20/50 or worse outcome (hazard
ratio = 0.43, 95% CI: 0.23, 0.81, P<0.01).
Conclusions: Ocular complications and vision loss were common in our cohort.
Active uveitis had a dose-dependent association with vision loss and use of
immunosuppressive drugs reduced the risk of vision loss; thus reinforcing the
recommendation that aggressive control of intraocular inflammation is critical to
the prevention of visual loss among patients with JIA-related uveitis.
Commercial Relationships: Jennifer E. Thorne, None; Anthony Gregory,
None; Ebenezer Daniel, None; C Stephen Foster, None; Douglas A. Jabs,
None; Grace A. Levy-Clarke, None; Robert B. Nussenblatt, None; James T.
Rosenbaum, None; Eric B. Suhler, None; John H. Kempen, None
Support: NIH/NEI R01 EY014943, Research to Prevent Blindness, Mackall
Foundation, NEI Intramural Funds, Dept of Veterans Affairs, KURE Fund
Program Number: 6254 Poster Board Number: D1114
Copyright 2012 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2012 Annual Meeting Abstracts
Presentation Time: 8:30 AM - 10:15 AM
In Search Of Intraocular Biomarkers In Uveitis Associated With Juvenile
Idiopathic Arthritis (jia)
Viera Kalinina Ayuso1A, Jolanda D. de Groot-Mijnes2, Jojanneke Dekkers1B,
Lenneke de Visser1A, Aniki Rothova1A, Joke H. de Boer1A. AOphthalmology,
B
Virology, 1UMC Utrecht, Utrecht, The Netherlands; 2Virology, Univ Medical
Center Utrecht, Utrecht, The Netherlands.
Purpose: To investigate the presence of specific protein profiles in aqueous
humour (AH) from patients with juvenile idiopathic arthritis (JIA) associated
uveitis.
Methods: Totally 116 children participated in this study. Aqueous humor (AH)
(n=73) and serum samples (n=105) were collected. The samples were analyzed
using surface-enhanced laser desorption/Ionisation time-of-flight (SELDI-TOF)
technique. The samples were divided in the following 4 groups: definitive JIA;
presumed JIA (not meeting the International League against Rheumatism criteria
completely); other uveitis entities and non-inflammatory controls.
Results: In the definitive JIA group significantly higher expression of five proteins
was found compared with other uveitis entities and controls (6672; 8725; 13762;
22313; 33410 Da). Definitive JIA and presumed JIA samples showed comparable
protein profiles in AH and did not differ in any of the peak cluster intensities.
Expression of protein peaks in AH showed significant positive and negative
bivariate correlations. Both definitive and presumed JIA samples could be
distinguished from other uveitis entities and controls by the presence of 13762 Da
protein and absence of 8255 Da protein in AH. Protein expression in AH of patients
with JIA and suspect JIA uveitis was strongly associated with activity of
inflammation.
Conclusions: Our results suggest the presence of intraocular JIA-specific proteins
which expression is correlated with activity of uveitis independently of definitive or
presumed JIA status. This could indicate similar molecular intraocular processes in
both groups of patients and support the hypothesis of an atypical manifestation of
JIA with primary uveitis. Further identification of potential biomarkers should be
performed.
Commercial Relationships: Viera Kalinina Ayuso, None; Jolanda D. de
Groot-Mijnes, None; Jojanneke Dekkers, None; Lenneke de Visser,
None; Aniki Rothova, None; Joke H. de Boer, None
Support: Dr. F.P. Fischer stichting, ODAS stichting, SNOO
Program Number: 6255 Poster Board Number: D1115
Presentation Time: 8:30 AM - 10:15 AM
Dramatic Effect Of Bolus Cyclophosphamide In Two Severe Cases Of Lupus
Retinopathy
Soydan Kurun, Andy Locatelli, Toufik Maalouf, Karine Angioi. Ophthalmology,
Regional Univ Hosp Ctr Brabois, Nancy, France.
Purpose: Reporting dramatic effect of bolus cyclophosphamide in two severe cases
of lupus retinopathy occurred in young women.
Methods: Two young women aged 22 and 31 with active systemic lupus
erythematosus (SLE) consulted for decreased visual acuity (4/10 P4 OU, and 8/10
P2 OD, 1/20 P14 OS respectively).
Results: In our two patients, fundus examination highlighted a bilateral macular
oedema, numerous cotton wool spots, perimacular exudates, flame hemorrhages
suggestive of the diagnosis of lupus retinopathy. Clinical and biological criteria
allowed the diagnosis of severe SLE due to neurolupus in one case, and lupus
glomerulonephritis in the other.
Aggressive treatment based on steroids and cyclophosphamide cures was decided
because of presence of both sight- threatening lesions and multi systemic
involvement.
Rapid improvement in systemic and biological signs of SLE was observed with
parallel disappearance of retinal signs. Full recovery of visual acuity was obtained
within 2 to 3 months in our two patients.
Prevalence of SLE varies between 10 and 50 p 100,000. Presence of
ophthalmological manifestations is estimated between 5 and 50% of the patients
depending on the series, and the most frequent one is lupus retinopathy. Prompt
treatment combining corticosteroids and cyclophophamide cures followed by
mycophenolate mofetil enables regression of retinal signs and improvement in
visual acuity, along with clinical and self- immune and inflammatory tests (C3, C4,
Anti Nuclear Factor and Anti DNA Antibodies)
Conclusions: In severe active forms of SLE an immediate specific treatment
combining cyclophosphamide cures and corticosteroids is mandatory to obtain not
only improvement of systemic manifestations but also complete visual recovery.
Commercial Relationships: Soydan Kurun, None; Andy Locatelli,
None; Toufik Maalouf, None; Karine Angioi, None
Support: None
Melissa Meyer zu Hörste1,2, Elena Stroeher1, Yang Zhang2, Katharina Roeck3,
Jens Fischer3, Utta Bercher-Pfannschmidt1, Anja K. Eckstein1, Erich Gulbins2.
1
Department of Ophthalmology, University Hospital Duisburg-Essen, Essen,
Germany; 2Department of Molecular Biology, University of Duisburg-Essen,
Essen, Germany; 3Institute of Pharmacology and Clinical Pharmacology,
University of Dueseldorf, Duesseldorf, Germany.
Purpose: To study mechanisms of pathologic proliferation of orbital fibroblasts
and the transformation of these cells to adipocytes, excessive production of
extracellular matrix, and inflammation which are hallmarks in the pathogenesis of
Graves ophthalmopathy (GO). These processes finally result in remodeling of the
orbital tissue with the typical symptoms of lid retraction, periorbital swelling,
disfiguring proptosis, optic nerve compression, and impaired ocular motility caused
by fibrotic changes in the extraocular muscles.
Methods: Human orbital fibroblasts were obtained from 12 patients with active,
severe GO and from 12 healthy control subjects. The cells were characterized by
immunofluorescence assay and flow cytometry. Expression of acid
sphingomyelinase (ASM) was determined by Western blot techniques, ASM
activity was measured by degradation of [14C]-labeled sphingomyelin to [14C]labeled phosphorylcholine, and Ceramide-levels were detected by DAG kinase
assay and immunofluorescence in orbital fibroblasts. Cell proliferation was
determined by BrdU-incorporation, hyaluronan (HA) production was assessed by a
HA-binding protein based assay, intracellular reactive oxygen species (ROS) were
determined by the dichlorofluorescein assay, and cell viability was measured by the
MTT-assay.
Results: ASM activity was inhibited with amitryptiline and with siRNA
technology. We demonstrate that a stable change in the phenotype of orbital
fibroblasts from GO patients results in a constitutive overactivity of ASM and a
high release of ceramide in these cells. High activity of the acid
sphingomyelinase/ceramide system in GO orbital fibroblasts results in a
constitutive increase of proliferation, release of reactive oxygen species, and
formation of hyaluronan. These hallmarks of GO are corrected upon pharmacologic
or siRNA-mediated inhibition of acid sphingomyelinase activity.
Conclusions: Our findings identify the acid sphingomyelinase/ceramide system as
a key mechanism in the pathophysiological changes affecting orbital fibroblasts
during GO and provide a rationale for inhibiting acid sphingomyelinase activity as
a novel treatment for GO.
Commercial Relationships: Melissa Meyer zu Hörste, None; Elena Stroeher,
None; Yang Zhang, None; Katharina Roeck, None; Jens Fischer, None; Utta
Bercher-Pfannschmidt, None; Anja K. Eckstein, None; Erich Gulbins, None
Support: IFORES grant Nr. D/D/107-40170
Program Number: 6256 Poster Board Number: D1116
Presentation Time: 8:30 AM - 10:15 AM
Inhibition Of The Acid Sphingomyelinase/ceramide System Prevents
Hallmarks Of Graves Ophthalmopathy
Copyright 2012 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Office at [email protected].