EBV-LMP2A肽诱导的特异性CTL对EBV阳性胃癌细胞的体外杀伤作用  

Cytotoxicity of specific CTL induced by EBV-LMP2A peptide on human EBV associated gastric carcinoma cell in vitro

作　　者：杜娟[1] 苏舒[1] 孟凡岩 邵洁[1] 徐秋萍[1] 邹征云[1] 刘宝瑞[1] 

机构地区：[1]南京大学医学院附属鼓楼医院肿瘤中心暨南京大学临床肿瘤研究所,江苏南京210008

出　　处：《中国肿瘤生物治疗杂志》2017年第9期938-943,共6页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.81602077;No.81572601)~~

摘　　要：目的:探讨HLA限制性EB病毒(Epstein-Barr virus,EBV)表位肽EBV-潜伏膜蛋白2A(EBV-latent membrane protein2A,EBV-LMP2A)诱导的细胞毒性T淋巴细胞(CTL)对EBV阳性胃癌细胞的体外杀伤作用。方法:选用南京大学医学院附属肿瘤医院肿瘤中心HLA-A2阳性胃癌患者外周血单个核细胞(PBMC)、人胃腺癌细胞株(AGS)和人EBV阳性胃腺癌细胞株(AGSEBV),通过改良的体外细胞培养技术用HLA-A2限制性的EBV-LMP2A抗原肽从人PBMC中诱导扩增出特异性CTL,采用四聚体技术和流式细胞术检测抗原肽诱导产生的特异性CTL的含量,通过FITC-PI标记流式术检测EBV-CTL对EBV阳性和EBV阴性的胃癌细胞的体外杀伤作用。结果:经改良的细胞培养技术和抗原肽双重刺激后EBV-LMP2A抗原肽可以诱导产生出高比例的抗原特异性CTL[EBV-LMP2A-356特异性T细胞占CD8+T细胞的(47.1±5.2)%];EBV-CTL对EBV阳性胃癌细胞的体外杀伤作用较EBV阴性的胃癌细胞明显增强[(45.1±9.3)%vs(19.4±2.5)%,P<0.05]。结论:通过改良的细胞培养技术使用EBV-LMP2A抗原肽能诱导产生高比例的EBV特异性CTL,其对EBV阳性胃癌细胞有较强的特异性杀伤作用。Objective：To study the cytotoxicity of specific cytotoxic T lymphocyte（CTL） induced by EBVLPM2 A peptide on human EBV associated gastric cancer cells in vitro.Methods：The peripheral blood mononuclear cells（PBMC） of HLA-A2 positive gastric cancer patients from Cancer Center of Drum Tower Hospital Affiliated to Medical School of Nanjing University,human gastric adenocarcinoma cell line（AGS） and human EBV positive adenocarcinoma cell line（AGS-EBV） were selected.By modified in vitro cell culture technique,specific CTL was amplified from human PBMCs with the induction of HLA-A2 restricted EBV-LMP2 A peptide;the content of specific CTL induced by peptide was detected by the method of tetramer and Flow cytometry;the in vitro cytotoxicity of EBV-CTL on both EBV＋and EBV-human gastric cancer cells was detected by FITC-PI.Results：Modified cell culture technique and EBV-LMP2 A peptide can induce a high proportion of antigen specific CTL（EBV-LMP2A-356 specific T cells accounted for [47.1±5.2]% of CD8＋T cells）;the cytotoxicity of EBV-CTL on EBV＋gastric cancer cells was significantly stronger than that of EBV-gastric cancer cells（[45.1±9.3]% vs [19.4±2.5]%,P〈0.05）.Conclusion：Using the modified cell culture technique,EBV-LMP2 A antigen peptide can induce a high proportion of EBV specific CTL,which has high specific cytotoxicity on EBV＋gastric cancer cells.

关 键 词：EB病毒-潜伏膜蛋白2A 细胞毒性T淋巴细胞 EB病毒阳性胃癌 细胞毒作用 

分 类 号：R730.51[医药卫生—肿瘤] R735.2[医药卫生—临床医学]