TCR-CDR3片段ASSLSGENTEAF在肝脏移植急性免疫排斥过程中的调控作用与机制研究

项目名称： TCR-CDR3片段ASSLSGENTEAF在肝脏移植急性免疫排斥过程中的调控作用与机制研究

项目编号： No.81500495

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 韩菲菲

作者单位： 首都医科大学

项目金额： 18万元

中文摘要： 肝脏移植是目前治疗肝脏衰竭的唯一有效手段，移植排斥的诊断与治疗是肝脏移植重要环节。肝脏移植排斥过程中T细胞发挥重要作用，T细胞受体（TCR）是T 细胞识别HLA/抗原肽复合物并与之结合从而发挥杀伤毒性的关键元件，对其高度可变区（CDR3）的研究能够让我们更加深入的认识移植排斥过程。本研究组前期应用arm-PCR 与高通量测序，首次发现在多数排斥个体中存在CDR3片段ASSLSGENTEAF，而在未发生免疫排斥和健康个体中没有检测到，因此表达此片段的T细胞克隆可能在免疫排斥反应中发挥重要作用。本研究进一步扩大样本量，在发生排斥的肝穿组织切片中验证表达ASSLSGENTEAF的T淋巴细胞特异性存在；采用体外混合淋巴细胞反应模型，研究封闭该TCR谱系后对淋巴细胞的激活情况影响及其作用机制，探讨是否能通过抑制排斥特异性TCR克隆而实现调控肝脏移植后发生的免疫排斥反应，为免疫排斥的治疗提供新的思路。

中文关键词： 肝移植；移植排斥；TCR-CDR3；混合淋巴细胞反应

英文摘要： Liver transplantation is currently the only effective mean for the treatment of liver failure. However, the change of T cell lineage drift was still unclear in the process of liver allograft rejection and immune tolerance. T cells are likely to have important roles in the process of liver allograft rejection. Analysis of the immune repertoire of T cell receptor (TCR), especially the CDR3 region is important for the determination of potential oligoclonal receptor skewing in a rejection population. In the preliminary studies we have found that there existed a TCR-CDR3 sequence by Arm-PCR and high throughput sequencing only in allograft rejection patients but not in stable patients and healthy volunteers. On the basis of the result we deduced that T cells expressing this clone might play an important role in the immune rejection. The project intends to predict the effects and mechanism of the specific TCR - CDR3 fragment ASSLSGENTEAF.. First, we intend to further expand the sample size and perform a dynamic monitoring in the process of rejection. Second, using mixed lymphocyte reaction model, investigate the effect of specific TCR - CDR3 by blocking it by specific antibody. Herein, we aimed at establishing and analyzing the TCR repertoire in liver allograft patients by means of multiplex PCR and CDR3-spectratyping with NGS to demonstrate any obvious CDR3 clonal expansions in patients with allograft as compared with control donors, which may help to provide an insight into the pathogenesis, diagnosis and treatment of the disease.

英文关键词： Liver transplant;allograft rejection;TCR-CDR3;mixed lymphocyte reaction