Molecular immunological investigation of Vogt-Koyanagi-Harada disease

建立了Vogt-Koyanai-Harada(VKH)病动物模型。该模型是用酪氨酸酶家族蛋白免疫诱导的，组织学结果与人类VKH病基本一致。我们用VKH病急性期和未治疗期患者的PBMC进行了抗酪氨酸酶家族蛋白的淋巴细胞增殖试验。这些淋巴细胞对酪氨酸酶和/或TRP1来源的一种或多种多肽表现出增殖反应。因此，我们提出VKH病的靶抗原是酪氨酸酶家族蛋白，并从YKH病患者的PBMC中建立了T细胞克隆。来自VKH患者的TCC对能与HLABRB1*0405相互作用的多肽具有反应性。这些TCC为Th1细胞，可能在VKH的发病中起重要作用。最后，我们从VKH患者的房水和脑脊液中建立了TCC。我们可以从VKH新鲜期和初治期患者中的7例建立TCC。建立了大量的TCC，并检测了30个克隆对酪氨酸酶家族蛋白的反应性，其中21个来自房水，9个来自脑脊液。令人惊讶的是，10/21的房水中的TCC和2/9的脑脊液中的TCC对酪氨酸酶或TRP1有增殖反应。这些比例非常高。我们现在正在用RT-PCR SSCP分析T细胞受体(TCR)。这些方法可能会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 histologie 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 taht the target antigens of VKH disease are tyrosinase family proteins.Then, we established T cell clone from PBMC of the YKH disease patients. The TCCs from VKH disease patients showed reactivity against the peptides that can interact with the HLA BRB1*0405. These TCCs were Th1 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 weretested 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.