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Analysis of TCR/HLA/ peptide interaction and investigation of etiology of autoimmune diseases

TCR/HLA/肽相互作用分析及自身免疫性疾病病因学研究

基本信息

批准号：
11694294
负责人：
NISHIMURA Yasuharu
金额：
$ 6.25万
依托单位：
KUMAMOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11694294/
关键词：
Autoimmune diseases Molecular mimicry Type I diabetes Vogt-Koyanagi-Harada disease Autoreactive CD4^+ T cells Expression cloning of T cell epitopes HLA class II T cell response 感染性微生物グルタミン酸脱炭酸酵素65 MHCクラスII分子発現クローニング Vogt-小柳-原田病 /

项目摘要

In the previous studies, we discovered assedations between specific HLA class II alleles and susceptibility to autoimmune diseases, and examined binding peptide motifs of disease-associated HLA class II molecules. In this study, to clarify the mechanisms of the disease-association of specific HLA alleles, we carried out the following researches. Identification of self peptides recognized by disease-related autoreactive T cell clones. We established many auto-antigen specific CD4^+ T cell clones from Asian type multiple sclerosis (MS) and anti-phosopholipid antibody syndrome (APS) patients. We identified many epitopes derived from self antigens and restricting HLA class II molecules, and examined the cytokines produced by the T cell clones. In addition, we searched for the target autoantigen of Vogt -Koyanagi-Harada disease (VKH), an autoimmune diseas e in which inflammatory disorders occur in multiple organs containing melanocytes. By SEREX method, we identified LEDGF (Lens Epithelial … More Cel l Derived Growth Factor) and UACA (Uveal Autoantigen with Coiled coil domains and Ankyrin repeats) as candidates for the target antigen. 2) Establishment of a method to identify cross-reactive epitopes recognized by disease-related autoreactive T cells.We established an expression cloning system to identify epitopes for CD4^+ T cell clones. Using this system, we examined the pattern of epitopes cross-recognized by two CD4^+ T cell clones established from Type I diabetes patients and specific to glutamate decarboxylase 65 (GAD65). Based on this information, we identified several mimicry epitopes of microbial antigen origin. 3) Analysis of T cell response induced by analogues of agonistic epilopes.We generated series of L cell transfectants expressing complexes of HLA-DR4 (DRB1^*0406) and agonist or partial agonist peptides in various densities. Using the transfectants, we discovered that a certain partial agonist peptide, when presented in a very high density, can induce T cell response in a similar magnitude as a full agonist peptide, andthat the T cell response is not accompanied by phosphorylation of ZAP-70 and LAT. We identified unique profiles of produced cytokines, down-modulation of TCR, and intracellular signaling events associated with this unique T cell response. Less

在之前的研究中，我们发现了特异性HLA II类等位基因与自身免疫性疾病易感性之间的关联，并检测了疾病相关HLA II类分子的结合肽基序。在本研究中，为了阐明特定HLA等位基因的疾病关联机制，我们进行了以下研究。疾病相关自身反应性T细胞克隆识别的自身肽的鉴定。我们从亚洲型多发性硬化症（MS）和抗磷脂抗体综合征（APS）患者中建立了许多自身抗原特异性CD4^+ T细胞克隆。我们从自身抗原和限制性HLA II类分子中鉴定了许多表位，并检测了T细胞克隆产生的细胞因子。此外，我们寻找Vogt -Koyanagi-Harada病（VKH）的靶自身抗原，VKH是一种自身免疫性疾病，炎症性疾病发生在含有黑素细胞的多个器官中。通过SEREX方法，我们鉴定出LEDGF（晶状体上皮细胞衍生生长因子）和UACA（具有卷曲线圈结构域和锚蛋白重复序列的葡萄膜自身抗原）作为候选靶抗原。2)建立疾病相关自身反应性T细胞识别的交叉反应性表位的鉴定方法。我们建立了一个表达克隆系统来鉴定CD4^+ T细胞克隆的表位。利用该系统，我们检测了来自1型糖尿病患者的两个CD4^+ T细胞克隆交叉识别的表位模式，这些克隆对谷氨酸脱羧酶65 （GAD65）具有特异性。基于这些信息，我们确定了几个微生物抗原来源的模仿表位。3)拮抗癫痫类似物诱导的T细胞应答分析。我们生成了一系列表达HLA-DR4 （DRB1^*0406）复合物和不同密度的激动剂或部分激动剂肽的L细胞转染物。使用这些转染物，我们发现，当以非常高的密度呈现时，某种部分激动剂肽可以诱导T细胞反应，其强度与完全激动剂肽相似，并且T细胞反应不伴随着ZAP-70和LAT的磷酸化。我们确定了产生的细胞因子的独特特征，TCR的下调，以及与这种独特的T细胞反应相关的细胞内信号事件。少