MICA反应性Vδ1γδT细胞对上皮肿瘤细胞的细胞毒作用

Cytotoxicity of MICA-reactive Vδ1γδT Cells towards Epithelial Tumor Cells

目的研究人类MHCⅠ类链相关基因A(MICA)是否可以在体外诱导Vδ1γδT细胞的扩增,并且对MICA反应性Vδ1γδT细胞的细胞毒作用进行初步研究。方法采用分子克隆技术原核表达并纯化MICA蛋白,使用重组MICA蛋白在体外诱导肿瘤组织中的Vδ1γδ肿瘤浸润T淋巴细胞(TILs),3-(4,5-二甲基噻唑-乙-YL)-2,5-二甲基-四唑溴盐(MTT)法检测MICA反应性Vδ1γδTILs的细胞毒活性。结果原核表达系统pET30表达并纯化了MICA;固相化的MICA蛋白可以诱导肿瘤组织中的Vδ1γδTILs在体外扩增,同时Vδ1γδTILs对MICA+的肿瘤细胞具有明显杀伤活性。结论MICA反应性Vδ1γδT细胞可能是肿瘤过继免疫治疗的新的候选效应细胞。

Objective To confirm whether human MHC classⅠchain-related A(MICA)induces the amplification of Vδ1γδtumor-infiltrating lymphocytes(TILs)in vitro and to identify the cytotoxicity of MICA-reactive Vδ1γδTILs towards epithelial tumor cells.Methods MICA protein was prokaryoticly expressed and purified by molecular cloning technology.The purified recombined MICA(rMICA)was used to induce Vδ1γδT cells from tumor tissues in vitro and the cytotoxicity of these Vδ1γδTILs were tested by3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT).Results The rMICA was expre-ssed in prokaryocyte with pET 30 as a vector.The immobilized rMICA protein could markedly induce the amplification of Vδ1γδT cells from tumor tissue in vitro.These Vδ1γδT cells showed strong cytolytic activities towards tumor cell lines expressing MICA.Conclusion The MICA-reactive Vδ1γδT cell may be a candidate for adoptive cellular therapy of tumors.