Immunological diseases of the eye (WS-099)Chairpersons: Koh-hei Sonoda, Nobuyuki Ohguro

International Immunology, Aug 2010

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Immunological diseases of the eye (WS-099)Chairpersons: Koh-hei Sonoda, Nobuyuki Ohguro

WS/PP-099-01 Amelioration of experimental autoimmune uveoretinitis by blockade of osteopontin with antibody or small interfering RNA K. Iwabuchi K. Iwabuchi 1 Div Immunobiol, Inst Genetic Med, Hokkaido University, Sapporo, Japan 2 Dept Ocular Inflamm Immunol, Grad Sch Med, Hokkaido University, Sapporo, Japan D. Iwata D. Iwata 3 Dept Ophthalmol, Grad Sch Med, Hokkaido University, Sapporo, Japan M. Kitamura M. Kitamura 3 Dept Ophthalmol, Grad Sch Med, Hokkaido University, Sapporo, Japan N. Kitaichi N. Kitaichi 3 Dept Ophthalmol, Grad Sch Med, Hokkaido University, Sapporo, Japan S. Kon S. Kon 4 Div Mol Immunol, Inst Genetic Med, Hokkaido University, Sapporo, Japan H. Kitamei H. Kitamei 3 Dept Ophthalmol, Grad Sch Med, Hokkaido University, Sapporo, Japan K. Namba K. Namba 3 Dept Ophthalmol, Grad Sch Med, Hokkaido University, Sapporo, Japan K. Yoshida K. Yoshida 3 Dept Ophthalmol, Grad Sch Med, Hokkaido University, Sapporo, Japan S. Ishida S. Ishida 3 Dept Ophthalmol, Grad Sch Med, Hokkaido University, Sapporo, Japan S. Ohno S. Ohno 2 Dept Ocular Inflamm Immunol, Grad Sch Med, Hokkaido University, Sapporo, Japan S. R. Rittling S. R. Rittling 5 The Forsyth Institute, Boston, MA, United States D. T. Denhardt D. T. Denhardt 6 Dept Cell Biol Neurosci, Rutgers University, Piscataway, NJ, United States T. Uede T. Uede 4 Div Mol Immunol, Inst Genetic Med, Hokkaido University, Sapporo, Japan K. Onoe K. Onoe 1 Div Immunobiol, Inst Genetic Med, Hokkaido University, Sapporo, Japan Abstract Experimental autoimmune uveoretinitis (EAU) in mice has been extensively studied as a model of human endogenous uveitis, which is common sight-threatening intraocular inflammatory disease. We have examined an aggravating role of osteopontin, a protein of pleiotropic functions and contributing to the development of Th1/Th17 cell-mediated immunity. Wild type (WT; C57BL/6) and OPN−/− mice were immunized with human interphotoreceptor retionoid-binding protein (hIRBP) peptide sequence 1-20 to induce EAU. OPN−/− mice showed milder disease progression compared with WT in clinical and histopathological EAU scores with reduced Ag-specific cell proliferation and pro-inflammatory cytokine production. OPN was elevated in WT mice along with the progression of EAU. Thus, OPN blockade was performed in WT mice with administration of either M5 antibody specific to SLAYGLR sequence, which is a cryptic binding site to integrins within OPN created by thrombin cleavage, or a small interfering RNA (siRNA) targeted on OPN coding sequence. Both treatments with either M5 antibody or siRNA could prevent EAU development with suppression of TNF-α, IFN-γ, and IL-17 production. Our results suggest that OPN represents a novel therapeutic target to control uveoretinitis. However, both blockades have been applied at the time of immunization and application after the onset was not as effective as the preventive regimen. Thus, we are further seeking the therapeutic application of OPN blockade for actual amelioration of EAU. WS/PP-099-02 Breakdown of immune privilege and spontaneous autoimmunity in mice expressing a retina-specific T cell receptor R. Horai R. Horai 1 NEI/NIH, Bethesda, MD, United States P. B. Silver P. B. Silver 1 NEI/NIH, Bethesda, MD, United States R. K. Agarwal R. K. Agarwal 1 NEI/NIH, Bethesda, MD, United States M. J. Mattapallil M. J. Mattapallil 1 NEI/NIH, Bethesda, MD, United States K. Natarajan K. Natarajan 2 NIAID/NIH, Bethesda, MD, United States P. Wang P. Wang 1 NEI/NIH, Bethesda, MD, United States C. Chan C. Chan 1 NEI/NIH, Bethesda, MD, United States R. R. Caspi R. R. Caspi 1 NEI/NIH, Bethesda, MD, United States Abstract Experimental autoimmune uveitis (EAU) is induced by immunization with retinal antigens and mimics human uveitis. We generated transgenic (Tg) mice that express a T cell receptor (TCR) specific to the uveitogenic retinal protein IRBP cloned from an autopathogenic (wild type) T cell line. The peripheral T cell repertoire of IRBP TCR Tg mice on the conventional background contained 15-30%, and on the RAG2-deficient background 90-95%, IRBP-specific CD4+ T cells, as judged by binding of Ag-specific MHC class II-Ig dimers. The cells proliferated to IRBP161-180 peptide and transferred EAU to naïve recipients. The proportion of memory T cells in the IRBP-specific population was substantially lower than in the polyclonal population, consistent with the notion of immune sequestration/privilege of the retinal antigens. Nevertheless, IRBP TCR Tg mice developed early spontaneous uveitis with most of the eye-infiltrating T cells displaying a memory phenotype. The ocular inflammatory infiltrate included Th1, Th17 and T regulatory cells (Tregs), detected by intracellular staining for IFN-γ, IL-17A, and Foxp3. IRBP-specific Tregs were enriched in the eye compared to the periphery. Interestingly, IRBP-specific Foxp3+ T cells did not seem to be positively selected in the thymus, suggesting that Tregs in the eye had been peripherally or locally converted from conventional T cells. We are currently studying where priming to “sequestered” retinal antigens occurs in this model. The IRBP TCR Tg mice provide a new valuable model of spontaneous uveitis to study basic mechanisms involved in maintenance and breakdown of immune homeostasis affecting privileged sites such as the eye. WS/PP-099-03 Common and different gene loci susceptible to sialoadenitis and dacryoadenitis in a Sjögren's syndrome mouse model T. Kamao T. Kamao Ehime University Graduate School of Medicine, Toon, Japan T. Miyazaki T. Miyazaki Ehime University Graduate School of Medicine, Toon, Japan Y. Soga Y. Soga Ehime University Graduate School of Medicine, Toon, Japan H. Komori H. Komori Ehime University Graduate School of Medicine, Toon, Japan M. Terada M. Terada Ehime University Graduate School of Medicine, Toon, Japan M. Nose M. Nose Ehime University Graduate School of Medicine, Toon, Japan Abstract Sjögren's syndrome (SS) is clinically manifested by sicca syndrome, associated with autoimmune sialoadenitis and dacryoadenitis. These lesions are commonly characterized by destruction of the ductules in the onset, associated with the accumulation of mononuclear cells, followed by destruction of the acinar glands. However, it still remains unclear whether these lesions develop coincidentally and in a common mechanism. An MRL/lpr strain of mice is well known as a SS mouse model, which spontaneously develops sialoadenitis and dacryoadenitis coincidentally. However, in the F2 generation of the intercrosses of MRL/lpr mice and non-SS-prone mice, C3H/lpr, these lesions were genetically dissociated. Thus, we graded the severity of both lesions in the F2 mice under histopathological scoring and then performed a quantitative trait locus analysis with polymorphic microsatellite markers. We identified three significant susceptibility loci to the severity of sialoadenitis on chromosomes 1 and 2, designated autoimmune sialoadenitis in MRL mice (Asm)3, Asm4 and Asm5. To those of dacryoadenitis, three significant susceptibility loci were determined on chromosomes 1, 2 and 5, designated autoimmune dacryoadenitis in MRL mice (Adacm)1, Adacm2 and Adacm3. Interestingly, Asm3 and Adacm1, and Asm5 and Adacm2 located in the common chromosomal regions each other, while Asm4 and Adacm3 were independent. These results indicate that sialoadenitis and dacryoadenitis may develop under the control of susceptibility genes with a different allelic combination to lead a regular variation of clinical phenotypes of SS. Positional candidate genes for the Adacm3 were discussed. WS/PP-099-04 Accumulation of regulatory T cells in the aqueous humour of uveitis patients S. H. Kottoor S. H. Kottoor 1 Institute of Biomedical Research, School of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom 2 Academic Unit of Ophthalmology, School of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom K. Morsley K. Morsley 1 Institute of Biomedical Research, School of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom 2 Academic Unit of Ophthalmology, School of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom R. Khanfer R. Khanfer 1 Institute of Biomedical Research, School of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom 2 Academic Unit of Ophthalmology, School of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom A. K. O. Denniston A. K. O. Denniston 1 Institute of Biomedical Research, School of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom 2 Academic Unit of Ophthalmology, School of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom I. J. Khan I. J. Khan 2 Academic Unit of Ophthalmology, School of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom K. S. Oswall K. S. Oswall 2 Academic Unit of Ophthalmology, School of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom M. Salmon M. Salmon 1 Institute of Biomedical Research, School of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom K. Piper K. Piper 3 School of Cancer Sciences, University of Birmingham, Birmingham, United Kingdom P. I. Murray P. I. Murray 2 Academic Unit of Ophthalmology, School of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom S. J. Curnow S. J. Curnow 1 Institute of Biomedical Research, School of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom 2 Academic Unit of Ophthalmology, School of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom Abstract The influx of leukocytes into the anterior chamber in uveitis is characterized by predominance of T lymphocytes which are assumed to play a role in the pathology. Accumulation of CD4+ regulatory T cells (Treg) at sites of inflammation has been shown in a number of autoimmune diseases. Here we wished to determine whether there are alterations in the number, phenotype or function of Treg in the aqueous humour (AqH) of uveitis patients. AqH (50-100µl) and peripheral blood were obtained from acute non infectious uveitis patients. Treg were defined as CD4+CD25highCD127low lymphocytes and were isolated from peripheral blood using Dynal beads or cell sorting. The frequency and expression of FoxP3 and CTLA-4 on Treg in uveitis AqH were significantly increased compared to peripheral blood (p<0.05). CD69, an early activation marker, was not expressed by these Treg, indicating that increased Foxp3 and CTLA-4 were not due to recent activation of conventional T cells. In vitro activation of peripheral blood Treg increased their FoxP3 expression to levels similar to those found on cells in the eye. Culture of Treg with the cytokines TGFβ2, TNF-α, IL-6, or IL-1β, which are present in uveitis AqH, had no effect on FoxP3 expression or Treg activity, and uveitis AqH did not affect the suppressive function of Treg in vitro. Our data suggest that Treg with a potent regulatory phenotype accumulate in the AqH of patients with uveitis. These cells may be actively suppressing the proliferation and function of pathogenic T cells, and therefore attempting to control inflammation. WS/PP-099-05 Pivotal roles of EBI3 for the initiation and maintenance of experimental autoimmune uveitis A. Takeda A. Takeda 1 Department of Ophthalmology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan 2 Deaprtment of Ophthalmology, National Kyushu Nedical Center, Fukuoka, Japan K. Sonoda K. Sonoda 1 Department of Ophthalmology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan E. Hasegawa E. Hasegawa 1 Department of Ophthalmology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan H. Yoshida H. Yoshida 3 Division of Molecular and Cellualr Immunoscience, Department of Biomolecular Sciences, Saga University, Saga, Japan T. Ishibashi T. Ishibashi 1 Department of Ophthalmology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan Abstract Epstein Barr induced gene 3 (EBI3), one of the components of IL-27, is a heterodimeric inflammatory cytokine. Recently, EBI3 also has been reported to be one of the components of IL-35 which mediates proliferation of CD4+CD25+ regulatory T cells. We have reported that IL-27 Receptor (WSX-1) plays important roles in Th1 induction in EAU. In this study, we investigated the role of EBI3 in experimental autoimmune uveitis (EAU). The clinical score of EAU in EBI3-deficient mice in the early phase was diminished as compared with that in WT. Consistently, histological analysis revealed that significant reduction of cellular infiltration into the retina was seen in EBI3 deficient mice. The induction of IFN-γ by draining lymph node cells from EBI3-deficient mice at day9 was less than that from WT, suggesting that EBI3 plays important roles in the induction of Th1. However, compared with WT mice, the score reached to the same level in EBI3-deficient mice at day22 and comparably diminished. The production of IL-17 by draining lymph node cells from EBI3-deficient mice at day9 was the same as that in WT, but IL-17 production in EBI3-deficient mice at day16 was increased as compared with that in WT. There was no difference of IL-10 and Foxp3 expression, which is concerned with CD4+CD25+ regulatory T cells, between EBI3-deficient mice and WT. These results indicate that EBI3 may affect Th1 response in the initiation phase and that EBI3 may suppress Th17 in the late phase independently of CD4+CD25+ regulatory T cells. © The Japanese Society for Immunology. 2010. All rights reserved. For permissions, please e-mail:


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Immunological diseases of the eye (WS-099)Chairpersons: Koh-hei Sonoda, Nobuyuki Ohguro, International Immunology, 2010, v21-v22, DOI: 10.1093/intimm/dxq358