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Analysis of T cell receptor repertoire in primary biliary cirrhosis

原发性胆汁性肝硬化T细胞受体库分析

基本信息

批准号：
05670477
负责人：
YAMAMOTO Kazuhide
金额：
$ 1.41万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05670477/
关键词：
Primary biliary cirrhosis T cell repertoire T cell receptor RT-PCR SSCP PCR SSCP PCR法

项目摘要

T cells play an important role in the destruction of bile duct in primary biliary cirrhosis (PBC). To analyze the T cells responsible for this bile ductt destruction, the T cell receptor (TCR) V beta repertoire was studied in liver biopsy specimens, and also in peripheral blood lymphocytes (PBL) obtained from 9 patients with PBC in the early stage (Scheuer's Stage I or II). The cDNA of each TCR V beta _<1-20> chain was prepared and amplified by reverse transcription-polymerase chain reaction (RT-PCR) and the PCR products were examined by single strand conformation polymorphism (SSCP) analysis. On the RT-PCR/SSCP analysis, a leukemic cell line, HPB-ALL,showed bands in TCR V beta 5.2 and V beta 6, indicating clonal expansion with distinct TCR.In PBL from healthy subjects, PCR products were amplified in most TCRV beta and were demonstrated as smears on SSCP,suggesting that PBL consists of diverse T cell clones. In PBC,most TCR V beta products were also amplified by RT-PCR in both liver tissues and PBL,and no biased expression of a particular V beta was observed. SSCP analysis revealed multiple bands in most V beta chains, suggesting the presence of selected but multiple T cell clones. Both the number and types of V beta showing clonal expansion were heterogeneous in PBC patients, although an increased number of clones was identified in V beta 6. These results suggest that T cells infiltrating the liver in PBC consist of multiple clonotypes and that T cells with TCR V beta 6 may play some role in the pathogenesis of PBC.

T细胞在原发性胆汁性肝硬化（PBC）中胆管的破坏中发挥重要作用。为了分析导致胆管破坏的 T 细胞，我们对肝活检标本以及从 9 名早期 PBC 患者（Scheuer Ⅰ期或 II 期）获得的外周血淋巴细胞 (PBL) 中的 T 细胞受体 (TCR) V β 谱进行了研究。通过逆转录聚合酶链式反应(RT-PCR)制备和扩增每个TCR Vβ_<1-20>链的cDNA，并通过单链构象多态性(SSCP)分析检查PCR产物。在 RT-PCR/SSCP 分析中，白血病细胞系 HPB-ALL 在 TCR V beta 5.2 和 V beta 6 中显示条带，表明具有不同 TCR 的克隆扩增。在来自健康受试者的 PBL 中，PCR 产物在大多数 TCRV beta 中扩增，并在 SSCP 上显示为涂片，表明 PBL 由不同的 T 细胞克隆组成。在PBC中，大多数TCR Vβ产物也在肝组织和PBL中通过RT-PCR扩增，并且没有观察到特定Vβ的偏向表达。 SSCP 分析显示大多数 V β 链中存在多个条带，表明存在选定的但多个 T 细胞克隆。尽管在 V beta 6 中发现克隆数量增加，但 PBC 患者中显示克隆扩张的 V beta 的数量和类型均存在异质性。这些结果表明，PBC 中浸润肝脏的 T 细胞由多种克隆型组成，并且带有 TCR V beta 6 的 T 细胞可能在 PBC 的发病机制中发挥一定作用。