Clonotype analysis of pathogenic T cells from patients with idiopathic thrombocytopenic purpura.

特发性血小板减少性紫癜患者致病性 T 细胞的克隆型分析。

• 批准号: 07671210
• 负责人: ODA Kenji
• 金额: $ 1.47万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671210/
• 关键词: thrombocytopenic purpura; T-cell receptor clonotype; single-strand conformation polymorphism; T-cell clone; autoimmune disease; 特発性血小板減少性紫斑病; T細胞クロノタイプ; SSCP解析

To determine whether clonal T cells accumulate in idiopathic thrombocytopenic purpura (ITP), we performed single-strand conformation polymorphism (SSCP) analysis to detect T-cell receptor (TCR) beta-chain usage of peripheral T cells. We detected significantly more oligoclonal T cells (15.5+/-8.9 band representative for clonal T-cell expansions) in peripheral blood from ITP patients than from healthy donors (2.8+/-2.6 bands). Frequently used V beta genes in these accumulated T cells in ITP were V beta 3,6,10,13.1 and 14. To determine whether these bands were derived from clonal T cells, presumably in a preactivated state, we established some T-cell clones (expressing CD4 and TCR V beta 6.13.1.or 14) by nonspecific stimulation from patients peripheral mononuclear cells, and examined their clonotypes. Clonal identities for three out of seven clones tested were confirmed using SSCP analyzes to compare the migration of their beta-chain complementarity determining region 3 (CDR3) cDNAs,expanded by polymerase chain reaction (PCR) with those from peripheral blood. Therefore, distinctive T-cell clones accumulated in the periphery in ITP and they may be related to the autoimmune-mediated distruction of platelets. We are currently trying to characterize those T-cell clones by identifying the T-cell antigens or epitopes and estimating the profile of their production of cytokines such as IL-4 and IFN gamma.

为了确定克隆性T细胞在特发性血小板减少性紫癜(ITP)中是否聚集，我们采用单链构象多态(SSCP)分析方法检测了外周血T细胞受体(TCR)β链的利用。我们在ITP患者外周血中检测到的寡克隆T细胞(代表克隆性T细胞扩增的15.5+/-8.9条带)明显多于健康献血员(2.8+/-2.6条)。在ITP中积累的T细胞中常用的Vβ基因是Vβ3、6、10、13.1和14。为了确定这些条带是否是来自克隆性T细胞，可能是处于预激活状态，我们从患者外周血单核细胞中通过非特异性刺激建立了一些表达CD4和TCR Vβ6.13.1或14的T细胞克隆，并检测了它们的克隆型。SSCP分析证实了7个克隆中的3个克隆的克隆性，以比较它们经聚合酶链式反应(PCR)扩增的β-链互补决定区3(CDR3)cDNAs的迁移性。因此，ITP患者外周血中可见特异性T细胞克隆聚集，可能与自身免疫介导的血小板破坏有关。我们目前正试图通过鉴定T细胞抗原或表位并估计它们产生IL-4和干扰素γ等细胞因子的情况来确定这些T细胞克隆的特征。

项目成果

藤村欣吾: "特発性血小板減少性紫斑病(ITP)におけるT細胞クロノタイプ解析" 厚生省特定疾患突発性造血障害調査研究班平成7年度研究業績報告書. 179-180 (1996)

Kingo Fujimura：“特发性血小板减少性紫癜（ITP）的T细胞克隆型分析”厚生省特发性造血疾病研究组FY1995研究成果报告179-180（1996）。

藤村欣吾: "特発性血小板減少性紫斑病(ITP)におけるT細胞クロノタイプ解析" 厚生省特定疾患特発性造血障害調査研究班 平成7年度研究業績報告書. 179-180 (1996)

Kingo Fujimura：“特发性血小板减少性紫癜（ITP）的T细胞克隆型分析”厚生省特发性造血障碍调查研究组1995财年研究成果报告179-180（1996）。

Fujimura, K et al: "Analysis of T-cell clonotype in Idiopathic thrombocytopenic purpura." 1995 Proc.Idiopathic Hematopoietic Disorders, Ministry of Health and Public Welfara Cooperation Work. 168-169 (1995)

Fujimura, K 等人：“特发性血小板减少性紫癜中 T 细胞克隆型的分析。”

下村壮司: "Oligoclonal accumulation of T cells in peripheral blood from patients with idiopathic thrombocytopenic purpura." British Journal of Haematology.95. 732-737 (1996)

Soji Shimomura：“特发性血小板减少性紫癜患者外周血中 T 细胞的寡克隆积累。”95，732-737（1996）。

Shimomura, T et al: "Oligoclonal accumulation of T cells in peripheral blood from patients with idiopathic thrombocytopenic purpura." British Journal of Haematology. 95. 732-737 (1996)

Shimomura, T 等人：“特发性血小板减少性紫癜患者外周血中 T 细胞的寡克隆积累。”