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

建立Vogt-Koyanai-Harada （VKH）病动物模型。该模型采用酪氨酸酶家族蛋白免疫诱导，其组织学表现与人VKH病基本一致。用PBMC对急性期和未治疗期VKH患者的酪氨酸酶家族蛋白进行淋巴细胞增殖试验。这些淋巴细胞对酪氨酸酶和/或TRP1衍生的一种或多种肽表现出增殖反应。由此，我们提出VKH病的靶抗原为酪氨酸酶家族蛋白。然后，我们从VKH患者的PBMC中建立T细胞克隆。来自VKH患者的tcc对可与HLA BRB1*04Q5相互作用的肽具有反应性。这些tcc是Thl细胞，可能在疾病的诱导中起重要作用。最后，我们从VKH患者的体液和脑脊液（CSF）中建立了tcc。我们可以从7例新鲜和未治疗期的VKH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个来自脑脊液）对酪氨酸酶家族蛋白的反应性。令人惊讶的是，10/21的房水tcc和2/9的脑脊液tcc对酪氨酸酶或TRP1表现出增殖反应。这些比率非常高。我们现在正在用RT-PCR SSCP分析T细胞受体（TCR）。这些方法可能揭示引起自身免疫性疾病的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.