肿瘤浸润性淋巴细胞亚群的功能研究及其受体克隆  

Functional Study and Cloning of Tumor Infiltrating Lymphocyte Subsets

作　　者：林晓为[1] 谢云青[1] 黄丽洁[1] 陈雪芳[1] 郑秋红[1] LIN Xiaowei;XIE Yunqing;HUANG Lijie(Fujian Cancer Hospital & Fujian Medical University Cancer Hospital,Fuzhou,350014)

机构地区：[1]福建省肿瘤医院,福建医科大学附属肿瘤医院福建省肿瘤生物治疗重点实验室,350014

出　　处：《实用癌症杂志》2020年第11期1749-1752,共4页The Practical Journal of Cancer

基　　金：国家临床重点专科建设资助项目;福建省引导性项目(编号:2018Y0015)。

摘　　要：目的比较肿瘤浸润性淋巴细胞(TIL)各亚群体外特异性杀伤肿瘤功能,摸索TIL细胞表面受体(TCR)互补决定区3(CDR3)基因体外扩增的方法,为寻找TIL细胞中肿瘤特异性抗原受体奠定前期实验基础。方法从肺癌肿瘤组织中分离出TIL细胞,利用CD8、PD-1抗体将TIL细胞分群,实时无标记细胞功能分析技术(real time cellular analysis,RTCA)分析比较各亚群细胞特异性杀伤活性。利用巢式RT-PCR的方法建立TCR VαCDR3区基因体外扩增的方法。结果TIL细胞中CD3^+CD8^+(CTL)细胞亚群占75.7%,PD-1的表达约为29.7%;CD8^+PD-1^+TIL细胞亚群比CD8^+PD-1-TIL细胞具有更强的体外杀伤原代肿瘤细胞功能;巢式RT-PCR成功扩增出TCR Vα1CDR3区基因片段。结论CD8^+PD-1^+TIL细胞亚群具有较强的特异性杀伤肿瘤活性;建立了通过巢式RT-PCR体外成功获取TCR所有CDR3区基因片段的实验条件。Objective To compare specific killing tumor effects of tumor infiltrating lymphocytes(TIL)subsets,and explore the experimental conditions of amplifiing the complementary determinant region 3(CDR3)gene of TIL cell surface receptor(TCR)in vitro to lay a foundation for searching informations of tumor specific antigens in TIL cells.Methods TIL cells were isolated from lung cancer tissues,and clustered by CD8 and PD-1 antibodies.Cell specific killing activity of each subgroup was analysis and compared by Real Time Cellular Analysis(RTCA).TCR Valpha CDR3 genes were amplified in vitro by nested RT-PCR.Results CD3+CD8+(CTL)subsets of TIL cells were accounted for 75.7%,and the expression of PD-1 was about 29.7%;CD8^+PD-1^+TIL subset has stronger killing effect on primary cancer cells in vitro than CD8^+PD-1-TIL;TCR Valpha1 CDR3 genes were successful amplified t by nested RT-PCR in vitro.Conclusion CD8^+PD-1^+TIL subset has stronger specific anti-tumor activity.The experimental conditions for obtaining all CDR3 gene from TIL TCR in vitro by nested RT-PCR were established.

关 键 词：肿瘤浸润性淋巴细胞 CD8 PD-1 T细胞受体 CDR3 

分 类 号：R730.21[医药卫生—肿瘤]