树突状细胞增强细胞因子诱导的杀伤细胞抗神经胶质瘤细胞作用及机制研究

Effects of dendritic cell enhanced cytokine induced killer cells against neuroglioma cells

目的探讨细胞因子诱导的杀伤细胞（CIK）与树突状细胞（DC）共培养后DC-CIK混合细胞抗神经胶质瘤细胞的免疫作用。方法分离健康人外周血单个核细胞，分别于体外诱导DC和CIK，然后共培养成DC-CIK细胞。实验分DC组、DC-CIK组、DC-T组和CIK组。Elisa试剂盒检测各组培养上清中IL-12和IFN．1的含量，流式细胞仪检测细胞表型，CCK一8法体外检测对神经胶质瘤细胞的杀伤活性。结果DC-CIK组培养上清中IL-12和IFN-1的含量分别为（110．24±2．22）mg／L和INF·Y／（913．46±20．64）mg／L，明显高于其它三组（P〈0．05）。DC—CIK组cDi细胞（61．34-1．31）％、CD3／CD56细胞（29．4±1．03）％也明显增加（P〈0．05）。对神经胶质瘤细胞的杀伤活性，DC—CIK组为（54．67±2．62）％，与DC组（19．44±1．07）％、DC—T组（21．27±1．85）％和CIK组（36．52±2．06）％比较，差异有统计学意义（P〈0．05）。结论DC—CIK细胞能诱导明显的神经胶质瘤细胞杀伤活性，为颅内肿瘤的免疫治疗提供了依据。

Objective To study the anti-neuroglioma immunifaction of DC-CIK cells from co-cul- ture of cytokine induced killer cells with dendritic cells. Methods Mononuclear cells were isolated from healthy human peripheral blood. DC and CIK were generated in vitro, and then cocultured together to pro- duce DC-CIK cells. The experiment was performed with four groups as DC group, DC-CIK group, DC-T group and CIK group. The level of IL-12 and IFN-~ in cultured supematant was detected by Elisa kit, and the phenotype was measured by flow cytometry（FCM）. The lymphocytes cytotoxic activity against neuro- glioma cell was analyzed by CCK-8 solution. Results The productions of IL-12 and IFN-~ in DC-CIK group were （ 110.24 -＋ 2.22）mg/L, （913.46 ＋ 20.64） mg/L separately, obviously higher than other three groups（P 〈 0.05 ）. Meanwhile, the phenotypes of CDs＋ （ 61.3 ± 1.31 ） % and CD3＋/CDs6 （ 29.4 ± 1.03 ） % were enhanced significantly （ P 〈 0.05 ）. DC-CIK cells induced higher cytotoxicity （ 54.67 ± 2.62） % against neuroglioma cells than that in DC group （ 19.44 ± 1.07 ） %, DC-T group （ 21.27 ± 1.85）% and CIK group（36.52 ±2.06）% （P 〈0.05）. Conclusion DC-CIK cells can induce potent immunifaction against neuroglioma, which provides experimental and theoretical basis for the immunothera- py for intracranial tumors.