基因扫描分析人T细胞受体CDR3谱型检测T细胞克隆技术的建立  

Established genescan technique of diversity gene of T cell antigen receptor CDR3 for assaying clonality of human TCR αβ T cell

作　　者：周健[1] 胡亮杉[1] 郭坤元[1] 黄迎[2] 郭爱林[2] 吴远彬[1] 王杨[1] 涂三芳[1] 

机构地区：[1]南方医科大学珠江医院,广东广州510282 [2]广东省人民医院

出　　处：《山东医药》2007年第32期7-9,共3页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(30471636)

摘　　要：目的建立人T细胞受体(TCR)互补决定区3(CDR3)基因谱型的基因扫描分析术,为分析T细胞克隆提供研究方法。方法分别以Jurkat和Raji细胞作为阳性和阴性对照,应用RT-PCR方法扩增5例健康志愿者外周血T细胞的34个TCR Vα亚家族和26个TCR Vβ亚家族的CDR3,并用基因扫描分析扩增产物以确定T细胞的克隆性。结果T细胞株Jurkat细胞仅表达TCR Vα1.1和Vβ8基因,且为单克隆,而B细胞株Raji细胞则不表达所有的TCR Vα和Vβ基因。5例健康人表达所有的TCR Vα和Vβ基因,均为多克隆T细胞。结论基因扫描分析人TCR CDR3基因谱型是检测T细胞克隆性的精确敏感方法。[ Objective] To establish genescan technique of diversity gene of T cell antigen receptor （TCR） complementary-determining region 3 （CDR3） for assaying clonality of human TCR αβ T cell. [ Methods] We took Jurkat cell and Raji cell as type strain and negative control respectively, and amplified 34 TCR α chain variable gene （TCR Vα） and TCR 26 β chain variable gene （TCR Vβ） of 5 healthy individuals peripheral blood T cell by RT-PCR. Productions of RT-PCR were assayed by genescan technique to determine clonality of TCR αβ T cell. [ Results] Only mRNA genes of TCR Vα 1.1 and TCR Vβ8 were found in Jurkat cells, no TCR Vet and TCR Vβ ge ne was found in Raji cells. Jurkat cell was monoclonal T cell. All the TCR Vα and TCR Vβ genes were expressed in 5 healthy individuals peripheral blood T cell. T cell were multiclonal. [ Conclusion] Genescan technique of diversity gene of TCR CDR3 was stable and precise method for assaying clonality of human TCR αβ T cell.

关 键 词：T细胞受体 互补决定区3 基因扫描 T细胞克隆 

分 类 号：Q343.1[生物学—遗传学]