流式细胞术检测CIK的免疫表型和体外杀伤活性

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。

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

Objective To investigate the immunophenotype and the cytotoxicity of cytokine-induced killer(CIK) against tumor cells in vitro.Methods Lymphocytes cells were isolated freshly from peripheral blood of healthy donors by Ficoll-Hypaque density centrifugation,and the cells obstained were induced by IFNγ,IL-2 and CD3McAb.Phenotypes and cytotoxicity of CIK were analysed by FACS.Target cells were differentiated from effect cells by CFSE dying.The mortality of Target cells were determined by FACS.Results The maximum proliferation of CIK reached 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 cytotoxicity against tumor cells of K562,HL-60,Hela,SMMC7721 and A375.Conclusion IFNγ,CD3McAb,IL-2 can induce peripheral lymphocytes to produce CIK which own strongly proliferation and exert highly efficient cytotoxic effects on tumor cells.