不同冻存时间对CIK细胞免疫表型及杀伤活性的影响  

Effects of different cryopreservation time on CIK cells immunophenotype and killing activities

作　　者：赵强[1] 豆兴芳 何苗[1] 张姝婷[1] 韩燕霞[1] 韩潇[1] 白海[1] 

机构地区：[1]兰州军区兰州总医院,甘肃兰州730050

出　　处：《现代肿瘤医学》2016年第10期1534-1536,共3页Journal of Modern Oncology

基　　金：甘肃省科技重大专项(编号:1102FKDA005)

摘　　要：目的:探讨不同冻存时间对外周血来源的CIK细胞免疫表型及其杀伤活性的影响。方法:采集6名健康患者外周血,分离外周血单个核细胞培养,第0h加入IFN-γ,24h后加入IL-2、抗CD3单抗,每隔3天补充含CD3单抗、IL-2的新鲜培养液,培养15天进行冻存。复苏冻存1、2、3、4、5月的CIK细胞为实验组,以常规培养至15天的CIK细胞为对照组。利用流式细胞仪检测各组CIK细胞中CD3^+CD4^+、CD3^+CD8^+、CD3^+CD56^+细胞的百分比,CCK8法检测各组CIK细胞对K562细胞的杀伤活性。结果:冻存1、2、3月的CIK细胞与对照相比CD3^+CD4^+、CD3^+CD8^+、CD3^+CD56^+细胞比例无明显差异(P>0.05),冻存4、5月的CIK细胞中CD3^+CD8^+、CD3^+CD56^+细胞比例较对照组明显下降[(70.62±6.35)%、(14.36±4.28)%、(67.87±5.63)%、(12.61±6.21)%vs(84.23±5.20)%、(22.75±3.46)%],P<0.05。对靶细胞的杀伤能力在1、2、3月时与对照相比未见明显减弱(P>0.05),冻存4月、5月后的CIK细胞在各效靶比对K562细胞的杀伤活性与对照相比明显减弱(P<0.05)。结论:冻存时间的长短对CIK细胞的免疫表型及杀伤活性均有一定的影响,且随着冻存时间的延长,CIK细胞的杀伤活性减弱。建议冻存CIK细胞的最佳时间为1、2和3月。Objective：To explore the effect of different cryopreservation time on CIK cells＇immunophenotype and killing activities. Methods ： PBMCs were obtained from 6 healthy adults, IFN - γ was added on day 0, anti - human CD3 and IL -2 were added after 24 hours, adding fresh culture medium containing CD3 monoclonal antibody and IL -2 were added according to the cell growth situation every 3 days, then continue culturing 15 days before cryopreservation. The cryopreserved CIK cells were reanimated monthly （ 1 - 5 months） , routine cultured CIK cells as control group. The proportion of CD3 ＋ CD4 ＋ , CD3 ＋ CD8 ＋ , CD3 ＋ CD56 ＋ cells were analyzed by flow cytometry. The cytotoxicity of CIK cells were tested by CCK8 experiment. Results：There was no significant difference in the proportion of CD3 ＋ CD4 ＋ , CD3 ＋ CD8 ＋ , CD3 ＋ CD56 ＋ cells between fresh CIK and cryopreserved CIK, however, the proportion of CD3 ＋ CD8 ＋ ,CD3 ＋ CD56 ＋ cells in cryopreserved CIK reanimated after 4 months, 5 months were lower than control group [ （70.62± 6.35 ） %, （ 14.36 ± 4.28） %, （67.87 ± 5.63 ） %, （ 12.61 ±6.21 ） % vs （ 84.23 ±5.20 ） %, （ 22.75 ± 3.46 ） % 1 , P 〈 0.05. The killing activity of cryopreserved CIK cells which reanimated after 4 months and 5 months at each effector - target ratio were weaker than control group （ P 〈 0.05 ）. Conclusion： The immunophenotype and killing activity of CIK cells were influenced by cryopreservation time, cryopreserve more than 3 months showed lower activity.

关 键 词：细胞因子诱导的杀伤细胞 冻存 免疫表型 杀伤活性 

分 类 号：R733[医药卫生—肿瘤]