流式细胞术检测CIK的免疫表型和体外杀伤活性  

Measurement of immunophenotype and cytotoxicity of CIK against tumor cells in vitro by FCM

作　　者：段秀梅[1] 谭岩[1] 宋燕[2] 王晓祺[1] 刘力华[1] 方艳秋[1] 许淑芬[1] 

机构地区：[1]吉林大学第一医院中心实验室,吉林长春130021 [2]吉林大学中日联谊医院普外科,吉林长春130033

出　　处：《吉林大学学报（医学版）》2006年第4期639-642,共4页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅重点科研项目资助课题(20050402-3);吉林省科技厅国际合作项目资助课题(20040707-2);高等学校博士学科点专项科研基金资助课题(20020183017)

摘　　要：目的:探讨细胞因子诱导的杀伤细胞(cytokine-induced killer,CIK)的免疫表型和体外杀伤活性。方法:从健康人外周血分离淋巴细胞,经过IFNγ、CD3McAb、IL-2诱导并培养,获得大量的CIK。采用流式细胞术检测CIK的表型;以CFSE标记靶细胞来区分效应细胞,再以PI染色,用FACS检测靶细胞的死亡率。结果:CIK于第22天达到增殖高峰,并高度表达CD3、CD11a、CD54和HLA-DR;中度表达CD3/CD56、CD25、CD28、CD69和FasL;CD16表达不增加。CIK对肿瘤细胞K562、HL-60、Hela、SMMC7721和A375均表现出杀伤活性,其中对K562、HL-60、Hela 3种细胞的杀伤作用较强;而对SMMC7721、A375的作用不明显。结论:细胞因子IFNγ、CD3McAb和IL-2体外诱导的单个核细胞具有强大的增殖力和杀伤活性。Objective To investigate the immunophenotype and the eytotoxieity of eytokine-indueed killer （CIK） against tumor eells in vitro. Methods Lymphoeytes eells were isolated freshly from peripheral blood of healthy donors by Fieoll-Hypaque density eentrifugation, and the eells obstained were indueed by IFNγ, IL-2 and CD3MeAb. Phenotypes and eytotoxieity of CIK were analysed by FACS. Target eells were differentiated from effeet eells by CFSE dying. The mortality of Target eells were determined by FACS. Results The maximum proliferation of CIK reaehed at the 22nd day. The phenotypes of CD3, CD11a, CD54, HLA-DR were expressed highly; CD25, CD28, CD69, FasL were expressed moderately on CIK. The expression of CD16 was not increased. CIK possessed the eytotoxieity against tumor eells of K562, HL-60, Hela, SMMC7721 and A375. Conclusion IFNT, CD3MeAb, IL-2 ean induee peripheral lymphoeytes to produee CIK whieh own strongly proliferation and exert highly effieient eytotoxie effects on tumor cells.

关 键 词：细胞因子诱导的杀伤细胞 免疫表型 杀伤活性 流式细胞术 

分 类 号：R392.12[医药卫生—免疫学]