IL-24基因修饰增强CIK细胞对HL-60细胞的细胞毒作用  

The cytotoxicity of IL-24 gene-modified CIK cells to HL-60 cells

作　　者：夏卫 郁心 王浦南 徐洪卫 陈宇 奚华新 杨吉成[2] 缪竞诚[2] 

机构地区：[1]无锡市红十字中心血站输血重点实验室,无锡214021 [2]苏州大学医学部基础医学与生物科学学院细胞与分子生物学教研室,苏州215123

出　　处：《中国免疫学杂志》2009年第12期1080-1084,共5页Chinese Journal of Immunology

基　　金：无锡市社会发展科技计划项目(CSZ00730)

摘　　要：目的:研究IL-24基因修饰的CIK细胞与同源树突状细胞共培养后对白血病细胞的杀伤作用及其机制。方法:从健康人外周血单个核细胞中常规诱导DC和CIK细胞,电穿孔法将IL-24基因导入CIK细胞中(获得细胞为CIK-IL24),RT-PCR和ELISA法检测CIK细胞中IL-24基因的表达,FCM和ELISA法检测转基因前后CIK表型及分泌细胞因子能力的变化,将CIK细胞和同源DC共培养,FCM法检测共培养的DC-CIK细胞对HL-60细胞细胞毒活性的变化。结果:通过电穿孔法成功将IL-24基因导入CIK细胞,与对照组相比,转IL-24基因后CIK细胞中CD3+、CD3+CD56+细胞的比例无明显改变,CD4+CD25+细胞比例显著下降。IL-24可上调CD3+CD56+细胞表面粘附分子CD54、CXCR4的表达,转染IL-24基因后CIK分泌TNF-α和IFN-γ的能力显著增强,与DC共同作用HL-60细胞时转染IL-24基因后的CIK细胞细胞毒活性明显增强。结论:通过IL-24基因修饰,明显增强了CIK细胞对HL-60细胞的杀伤能力,其机制与IL-24促进CIK分泌TNF-αI、FN-γ,上调CIK细胞表面粘附分子的表达,减少CD4+CD25+调节性T细胞比例等密切相关。Objective：To study the antitumor effect and mechanism of cocultured CIK cells modified with IL-24 gene and autologous DCs on HL-60 cells in vitro. Methods：DCs and CIK cells were prepared routinely from human peripheral blood mononuclear cells （ PBMC）. IL-24 gene was transferred into CIK cells via electroporation. The cells obtained were named CIK-IL24. RT-PCR and ELISA were used to evaluate expression of IL-24 gene in transfected CIK cells. The phenotypic changes of CIK cells were identified by llowcytometry analysis. The concentration of IFN-γ and TNF-α in supernatant of CIK was determined by ELISA. FCM was used to determine the cytotoxicity of cocultured CIK cells modified with IL-24 gene and autologous DCs against HL-60 cells. Results： Eukaryotic expressing plasmid pcDNA3.0-IL24 was transferred into CIK cells successfully via electroporation. The expressing rate of CD3 ^＋ . CD3^＋ CD56^＋ cells had no significant change in CIK cells. However, the rate of CD4^＋ CD25^＋ cells was significantly decreased compared with that of the control group. Expression of adhesion molecules CD54, CXCR4 were significantly increased on CD3^＋ CD56^＋ cells. CIK-IL24 cells produced markedly higher levels of IFN-γ and TNF-α as compared with the CIK cells. By comparison with non-transfected CIK cells co-cultured with DCs, transfected CIK cells co-cultured with DCs had a significantly higher lytic activity against HL-60 cells. Conclusion： IL-24 gene modification can enhance the anti-tumoral immunity of CIK cells, the mechanism of which might be related to the increased secretion of IFN-γ,TNF-α, up-regulation of adhesion molecule expression, and reduction of the rate of CD4^＋ CD25^＋ cells in CIK cells.

关 键 词：人IL-24 CIK细胞 HL-60细胞 细胞毒 

分 类 号：R392.1[医药卫生—免疫学]