猪瘟C株疫苗毒感染免疫特异的CD4+/CD8+ T细胞亚群αβTCR的分子结构特征及其免疫学功能

T细胞受体（αβTCR）在机体获得性细胞免疫中扮演着重要角色，但由于其基因的遗传多样性和结构复杂性，基于病毒感染或免疫特异的猪αβT细胞克隆化增殖研究在国际上尚属空白。项目针对猪瘟C株疫苗毒感染小型猪可激发其αβTCR胚系分化和基因重排而引起的T细胞克隆化增殖现象，采用流式细胞仪法分选和鉴定猪CD4+/CD8+ T细胞亚群，应用接头连接介导的通用PCR法高通量扩增、克隆和建立包含大量未知αβ链的TCR基因库；采用生物信息学技术分析αβTCR基因的分子结构特征及其多样性与特异性，探讨猪αβT细胞克隆化增殖规律；选择病毒感染免疫特异的CD4+/CD8+ T细胞的αβTCR分子，分别构建其荧光标记的动物细胞表达载体，转基因修饰特定CD4+/CD8+ T细胞后，研究其与靶细胞互作产生的免疫学效应，阐明其遗传多样性与免疫学功能特异性的分子基础，为猪瘟表位肽高效疫苗研究及其疫病控制提供理论依据。

αβT cell receptor(αβTCR) take an important role in host acquired cellular immunity.But investigation of T cell cloning proliferation in porcine infected by viruses is still not report due to the diversity and complexity of αβTCR.This project firstly sorting and identify the CD4+/CD8+ T cell subgroup with flow cytometry,and amplify,clone and establish the repertoires of αβ TCR gene by AL-PCR according the phenomenon of T cell cloning proliferation through the generearrangement of TCR stimulating by infection of C strain-swine fever virus.Then analyze the genetic constitution,structural feature and its diversity and specifity of αβTCR by technique of bioinformation,and highlight the rule of T cell cloning proliferation in infection. Thirdly,construct these recommbinant expression vector of TCR expressing in animal cell using information of the α or β TCR of CD4+/CD8+ T cell respectively, and transgenic modify defined CD4+/CD8+ T cell of porcine.And,observe and analyze immunological effect of interaction between the T cells and APCs.