Development of disease-specific diagnosis and immunotherapy for Sjogren's syndrome

干燥综合征的疾病特异性诊断和免疫治疗的发展

基本信息

批准号：
12557022
负责人：
HAYASHI Yoshio
金额：
$ 8.51万
依托单位：
The University of Tokushima
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557022/
关键词：
autoimmune disease Sjogren's syndrome autoantigen specific diagnostic system ELISA assay α-fodrin cDNA fusion protein peptide-based immunotherapy 抗原エピトープ高感度ELISA α-フォドリン疾患モデル NFS sldマウス

项目摘要

Previously we have identified a 120 kD α-fodrin as a common autoantigen in the pathogenesis of primary Sjogren's syndrome (SS) in humans, but the mechanisms underlying the immunodominant epitope recognition remain unclear. SS in human is a T cell-mediated autoimmune disease in the salivary and lacrimal glands, leading to clinical symptoms of dryness of the mouth and eyes (sicca syndrome). Recombinant α-fodrin protein, the cDNA encoding human α-fodrin (JS-1) was constructed by inserting cDNA into EcoR1 site of pGEX-2T, To establish disease-specific diagnostic system, approximately 200 sera from patients with SS were tested with ELISA assay using JS-1 recombinant protein. Autoreactive T cell clones that recognize synthetic N-terminal portion of α-fodrin autopeptide were established, which produced Th1 cytokines, and showed cytotoxic activities. Alanine scanning mutagenesis of the epitope peptide indicate that the two amino acid substitutions of MHC contact points within epitope are sufficient to alter peptide binding and MHC restriction. Moreover, it is evident that only two TCR contact points are essential for initiation of antigen-specific T cell response. An analogue peptide-based vaccination prevented the disease through downregulation of Th1 responses and autoantibody production. Blockade of pathogenic T cell activity through an analogue peptide-based immunotherapy is highly effective for T cell-mediated autoimmune disease such as human SS.

以前，我们已经将120kDα-佛教蛋白鉴定为人类原发性Sjogren综合征（SS）的常见自身抗原，但是在人类中，尚不清楚免疫主流表位识别的机制尚不清楚。人类中的SS是唾液和泪腺中T细胞介导的自身免疫性疾病，导致口腔和眼睛干燥的临床症状（Sicca综合征）。通过将cDNA插入PGEX-2T的ECOR1位点构建以建立疾病特异性诊断系统，使用JS-1重新组合蛋白测试了大约200名来自SS的患者的疾病特异性诊断系统，以建立疾病特异性的诊断系统来建立大约200名Sera，来建立大约200名疾病特异性诊断系统，从而构建了编码人α-峰蛋白（JS-1）的重组α-速蛋白蛋白（JS-1）的cDNA。建立了识别α-佛教蛋白自肽的合成N末端部分的自动反应性T细胞克隆，从而产生了Th1细胞因子，并显示了细胞毒性活性。表位肽的丙氨酸扫描诱变表明，表位中MHC接触点的两个氨基酸取代足以改变肽结合和MHC限制。此外，有证据表明，只有两个TCR接触点对于启动抗原特异性T细胞反应至关重要。基于模拟肽的疫苗通过下调TH1反应和自身抗体产生来阻止该疾病。通过基于模拟肽的免疫疗法阻断致病性T细胞活性对T细胞介导的自身免疫性疾病（例如人类SS）非常有效。