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

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.