CAJ | 학술논문

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-α、IFN-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.
목적:연구IL-24기인수식적CIK세포여동원수돌상세포공배양후대백혈병세포적살상작용급기궤제.방법:종건강인외주혈단개핵세포중상규유도DC화CIK 세포,전천공법장IL-24기인도입CIK세포중(획득세포위CIK-IL24),RT-PCR 화ELISA법검측CIK세포중IL-24기인적표체,FCM화ELISA법검측전기인전후CIK표형급분비세포인자능력적변화,장CIK 세포화동원DC공배양,FCM법검측공배양적DC-CIK세포대HL-60세포세포독활성적변화.결과:통과전천공법성공장IL-24기인도입CIK세포,여대조조상비,전IL-24기인후CIK세포중CD3~+、CD3~+CD56~+세포적비례무명현개변,CD4~+CD25~+세포비례현저하강.IL-24가상조CD3+CD56+세포표면점부분자CD54、CXCR4적표체,전염IL-24기인후CIK분비TNF-α화IFN-γ적능력현저증강,여DC공동작용HL-60세포시전염IL-24기인후적CIK세포세포독활성명현증강.결론:통과IL-24기인수식,명현증강료CIK세포대HL-60세포적살상능력,기궤제여IL-24촉진CIK분비TNF-α、IFN-γ,상조CIK세포표면점부분자적표체,감소CD4~+CD25~+조절성T세포비례등밀절상관.
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 flowcytometry 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.