A method to determine antigen peptides for the conally expanded T cells in systemic autoimmune diseases

系统性自身免疫性疾病中圆锥扩增 T 细胞抗原肽的测定方法

• 批准号: 11670465
• 负责人: KATO Tomohiro
• 金额: $ 2.5万
• 依托单位: St.Marianna University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670465/
• 关键词: systemic autoimmune disease; systemic lupus erythematosus; rheumatoid arthritis; MHC; T cell; antigen peptide; phage; peptide library

We previously reported oligoclonal T cell accumulation in the systemic autoimmune diseases. For instance, the T cell clonal expansion was detected in peripheral blood and also in affected kidney in systemic lupus erythematosus (SLE) and in synovial tissue in rheumatoid arthritis (RA). However, target molecules of these accumulated or expanded T cells remains to be determined, even though they are of great importance for understanding the pathogenesis of the diseases. To determine the antigen-peptides of the T cell clonotypes, we tried to establish screening system of antigenic peptides using random peptide phage library with MHC molecules and reconstructed TCR from clinical samples. As a result, we demonstrated that HLAB5101 and H-2Db with betamicroglobulin is expressed on phage with ability to bind antigenic peptides. Further, we obtained TCR a and b chain in pair from one T cell of clinical samples from patients with SLE, using single cell PCR and reconstructed the TCR as a single chain in vitro. Combination of the reconstructed TCR and randomized peptide library would be a powerful tool to determine the antigens for the clonally expanded T cells.

我们先前报道了寡克隆T细胞在系统性自身免疫性疾病中的积聚。例如，在外周血中检测到T细胞克隆性扩增，并且在系统性红斑狼疮（SLE）的受影响的肾脏中以及在类风湿性关节炎（RA）的滑膜组织中也检测到T细胞克隆性扩增。然而，这些积累或扩增的T细胞的靶分子仍有待确定，即使它们对于理解疾病的发病机制非常重要。为了确定T细胞克隆型的抗原肽，我们尝试利用随机肽库和MHC分子建立抗原肽筛选系统，并从临床样品中重建TCR。结果，我们证明了HLAB 5101和H-2Db与β-微球蛋白在噬菌体上表达，具有结合抗原肽的能力。此外，我们使用单细胞PCR从SLE患者的临床样品的一个T细胞中成对地获得TCR a和B链，并在体外将TCR重建为单链。重组TCR与随机肽库的结合将成为克隆扩增T细胞抗原鉴定的有力工具。

项目成果

Kato. T et al: "Comparison of T Cell receptor JB gene usage in spleen cells of different mouse strains"Microbiol Immunol. 43. 93-97 (1999)

加藤。

Yu X et al: "Anti-CD69 autoantibodies cross react with low density lipoprotein receptor-related protein 2 in systemic autoimmune diseases."J Immunol. 166・2. 1360-1369 (2001)

Yu X 等人：“抗 CD69 自身抗体与系统性自身免疫性疾病中的低密度脂蛋白受体相关蛋白 2 发生交叉反应”，《免疫学杂志》166·2（2001）。

Matsui. T et. al: "Autoantibodies to Tcell costimulatory molecules in systemic atuoimmune diseases"The Journal of Immunology. 162. 4328-4335 (1999)

松井。

Kato T et al: "Analysis of accumulated T cell clonotypes in patients with systemic lupus erythematosus."Arthritis Rheum. (In press).

Kato T 等人：“系统性红斑狼疮患者中累积的 T 细胞克隆型分析。”关节炎大黄。

Kurokawa M, T Jiankun, Matsui T, Masuko-Hongo M, Yabe T, Nishioka K, Yamamoto K, Kato T: "Paired-cloning of the T cell receptor α and β genes from a single cell without the establishment of a T cell clone."Clin Exp Immunol. 123(2). 340-345 (2001)

Kurokawa M、Tjiankun、Matsui T、Masuko-hongo M、Yabe T、Nishioka K、Yamamoto K、Kato T：“在不建立 T 细胞的情况下，从单个细胞配对克隆 T 细胞受体 α 和 β 基因克隆。”《临床实验免疫学》123(2)。340-345(2001)