CD_(80)基因导入人卵巢癌细胞系及其体外诱导细胞毒T淋巴细胞的研究  

Gene transfer of CD_( 80)into a human ovarian cancer cell line and induction of cytotoxic T lymphocyte in vitro

作　　者：崔保霞[1] 孔北华[1] 姜洁[1] 江森[1] 

机构地区：[1]山东大学齐鲁医院妇产科,济南250012

出　　处：《中华妇产科杂志》2001年第4期229-232,共4页Chinese Journal of Obstetrics and Gynecology

摘　　要：目的　用逆转录病毒载体将CD80 基因导入人卵巢癌细胞系TYK ,观察表达CD80 基因的TYK(TYK hCD80 )细胞体外诱导细胞毒T淋巴细胞 (CTL)的增殖及其杀瘤活性。方法　用逆转录病毒载体将CD80 基因导入TYK细胞 ,流式细胞仪测定其CD80 基因的表达。用TYK hCD80 细胞在抗CD3 单克隆抗体 (单抗 )存在下刺激外周血单个核细胞 (PBMC) ,诱导CTL ,3 H 胸腺嘧啶核苷掺入法测定其增殖活性 ,四甲基偶氮唑蓝法测定CTL的杀瘤活性。结果　转染后经G418(geneticin ,是一种氨基糖甙类抗生素 )筛选 ,流式细胞仪检测 ,CD80 基因表达率最高为 84 9%。TYK hCD80 、TYK在抗CD3 单抗存在下刺激PBMC增殖时 ,3 H 胸腺嘧啶核苷掺入量分别为 (4 0 6 0 4± 842 )、(80 0 0± 5 94)cpm ,两者比较 ,差异有显著性 (P <0 0 5 )。TYK hCD80 体外诱导的CTL较TYK诱导者对TYK细胞的杀伤率显著增高(P <0 0 5 )。结论　表达CD80 基因的卵巢癌细胞在抗CD3 单抗存在下能诱导产生CTL ,增殖快且有较高的杀伤活性 。Objective To investigate the proliferation and cytotoxicity of cytotoxic T lymphocyte (CTL) induced by ovarian cancer cells transfected with a CD 80 gene containing retroviral vector Methods The ovarian cancer cell line TYK cells were transfected with retro virus vector PLXSN hCD 80 After geneticin (G418) selection, the CD 80 expression of the transfectants was confirmed by flow cytometry CTL was induced by co culture of CD 80 expressing TYK cells (TYK hCD 80 ) and peripheral blood mononuclear cells (PBMC) in the presence of anti CD 3 monoclonal antibody (McAb) Proliferation of PBMC was measured using 3?H Thymidine incorporation assay The lysis activity of CTL toward tumor cells was determined using methyl thiazolyl tetrazolium (MTT) assay Results After transfection and G418 selection, the CD 80 expression was proved by flow cytometry The highest rate of CD 80 expression was 84 9% The TYK cell line expressing high CD 80 was named TYK hCD 80 In the presence of anti CD 3 McAb,TYK hCD 80 significantly enhanced proliferation of PBMC than TYK cells ( 3H Thymidine incorporation, (40 604± 842) vs (8 000± 594) cpm ( P <0 05) The lysis activity of CTL activated by TYK hCD 80 was higher than that of TYK( P <0 05) Conclusions Ovarian cancer cells expressing CD 80 can induce the production of CTLs which have high lysis activity and high proliferation rate in the presence of anti CD 3 McAb This study may provide the experimental basis for immunogenic therapy of ovarian cancer

关 键 词：卵巢肿瘤 CD80抗原 基因转染 T淋巴细胞 细胞毒性 

分 类 号：R737.31[医药卫生—肿瘤]