流式细胞术检测CIK的免疫表型和体外杀伤活性

doi:吉林省科技厅重点科研项目资助课题（200504

流式细胞术检测CIK的免疫表型和体外杀伤活性

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

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

收稿日期:2006-01-11 修回日期:1900-01-01 出版日期:2006-07-28 发布日期:2006-07-28
通讯作者: 谭岩

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

DUAN Xiu-mei¹, TAN Yan^1*, SONG Yan²，WANG Xiao-qi¹，LIU Li-hua¹，FANG Yan-qiu¹，XU Shu-fen¹

1. Central Laboratory, First Hospital, Jilin university, Changchun　130021,China; 2. Department of General Surgery,　China-Japan Union Hospital,Changchun　130033,China

Received:2006-01-11 Revised:1900-01-01 Online:2006-07-28 Published:2006-07-28
Contact: TAN Yan

摘要/Abstract

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。

关键词: 免疫表型, 杀伤活性, 流式细胞术

Abstract: 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,SMM C7721 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.

Key words: immunophenotype, cytotoxicity, flow cytometry

中图分类号:

R392.12

段秀梅，谭岩，宋燕，王晓祺，刘力华，方艳秋，许淑芬. 流式细胞术检测CIK的免疫表型和体外杀伤活性[J]. J4, 2006, 32(4): 639-642.

DUAN Xiu-mei, TAN Yan, SONG Yan，WANG Xiao-qi，LIU Li-hua，FANG Yan-qiu，XU Shu-fen. Measurement of immunophenotype and cytotoxicityof CIK against tumor cells in vitro by FCM[J]. J4, 2006, 32(4): 639-642.

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