Molecular immunological investigation of Vogt-Koyanagi-Harada disease

Vogt-小柳-原田病的分子免疫学研究

• 批准号: 13671818
• 负责人: YAMAKI Kunihiko
• 金额: $ 2.24万
• 依托单位: AKTTA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671818/
• 关键词: T cell clone; Tyrosinase family proteins; VKH disease; animal model; Tyrosinase TRP1; CD4 T cell(Th1 cell); TCRCT cell receptor

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.

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，其病理组织学表现与人VKH病基本一致，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YKH病患者的外周血单个核细胞中建立了T细胞克隆。来自VKH疾病患者的TCC显示出对可与HLA BRB 1 *0405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例新鲜和未经治疗的VKH病患者中建立TCC。建立了多个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CSF。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。

项目成果

Yamaki K: "Establishment and characterization of the T cell clones from the aqueous humons and cerebrospinal fluids with vogt-koyanagi-Harada disease patients"Akita. J. Med. (in press).

Yamaki K：“从 vogt-koyanagi-Harada 病患者的房水和脑脊液中建立 T 细胞克隆并对其进行表征”秋田。