Molecular immunological investigation of animal model of Vogt-Koyanagi-Harada disease

建立了Vogt-Koyanai-Harada（VKH）病动物模型。该模型由酪氨酸酶家族蛋白免疫诱导，其病理学表现与人VKH病几乎相同，我们用VKH病急性期和未治疗期的PBMC进行了抗酪氨酸酶家族蛋白的淋巴细胞增殖试验。这些淋巴细胞显示出对一种或多种衍生自酪氨酸酶和/或TRP 1的肽的增殖反应。由此我们提出VKH病的靶抗原是酪氨酸酶家族蛋白，并从VKH病患者PBMC中建立了T细胞克隆。来自VKH疾病患者的TCC显示出对可与HLA BRB 1 * 04 Q5相互作用的肽的反应性。最后，我们从VKH病患者的房水和脑脊液中建立了TCCs。我们可以从7例新鲜的、未经治疗的VKH病患者中建立TCC。建立了许多TCC，并测试了30个克隆（21个来自房水，9个来自CSF）对酪氨酸酶家族蛋白的反应性。令人惊讶的是，10/21的来自房水的TCC和2/9的来自CSF的TCC显示出对酪氨酸酶或TRP 1的增殖反应。这个比例非常高。我们正在用RT-PCR SSCP分析T细胞受体（TCR）。这些方法可以揭示导致自身免疫性疾病的T细胞。

We had established animal model of Vogt-Koyanai-Harada (VKH) disease. This model was induced by immunization of tyrosinase family proteins and the histologic findings of this disease was almost identical to that of human VKH disease.We had done lymphocyte proliferation assay against tyrosinase family proteins using PBMC of acute and untreated stage of VKH disease patients. These lymphocytes showed proliferative response against one or more of the peptides derived from tyrosinase and/or TRP1. From these facts, we proposed that the target antigens of VKH disease are tyrosinase family proteins.Then, we established T cell clone from PBMC of the VKH disease patients. The TCCs from VKH disease patients showed reactivity against the peptides that can interact with the HLA BRB1*04Q5. These TCCs were Thl cells and may play important role to the induction of the disease.Finally, we established TCCs from aqueous humors and cerebrospinal fluid (CSF) of VKH disease patients. We could establish TCCs from 7 of the fresh, and untreated stage of VKH disease patients. Many TCCs were established and 30 clones, 21 from aqueous humor and 9 from CSF were tested the reactivity against tyrosinase family proteins. Surprisingly, 10/21 of TCCs from aqueous humor and 2/9 of TCCs from CSF showed proliferative response against tyrosinase or TRP1. These ratios were very high. We are now analyzing the T cell receptor (TCR) with RT-PCR SSCP. These methods may reveal the T cells that cause the autoimmune disease.