IFN-γ与IL-12诱导CIK细胞生物学活性的比较研究

Comparative study on the biological features of cytokine-induced killer cells induced by IFN-γ and IL-12

探讨IFN-γ与IL-12分别对细胞因子诱导的杀伤细胞(cytokine-induced killer,CIK)的体外扩增速度、细胞免疫表型和杀伤活性的影响。分别以传统的IFN-γ、CD3 mAb、IL-2和IL-1α以及IL-12、CD3 mAb、IL-2和IL-1α为诱导剂制备CIK细胞,直接活细胞计数法比较两者的扩增速度,流式细胞仪检测比较两组细胞的免疫表型,LDH法比较两者对K562细胞株的杀伤活性。结果:两种CIK细胞的增殖速度无显著性差异,但IL-12启动的CIK细胞体外扩增时存活时间比常规CIK细胞长,并且其CD3+CD8+及CD3+CD56+细胞显著高于常规CIK细胞(P<0.05),CD3+CD4+和CD4/CD8显著低于常规CIK(P<0.05),并且其体外对K562的非特异杀伤活性在效靶比为40∶1时显著高于常规CIK。IL-12诱导扩增的CIK中效应细胞比例显著增加,同时抑制性T细胞比例降低,且体外该CIK细胞的杀伤能力较常规CIK好。故IL-12取代IFN-γ对CIK进行预刺激可增强CIK细胞的生物学活性。

To compare the proliferation,phenotypes and cytotoxicity of cytokine-induced killer cells（CIK） induced by IFN-γ and IL-12,these cells were stimulated with IFN-γ and IL-12 separately and followed by CD3mAb,IL-1α and IL-2.The proliferation of these cells was compared by cell counting,the phenotypes of CIK cells were analyzed by flow cytometry and the cytotoxic activities to K562 cells were determined by LDH assay.It was demonstrated that there was no significant difference between CIK induced by IFN-γ and IL-12 in the proliferation rate.However,the survival time of the in vitro proliferation of these two kinds of CIK cells was longer than that of the routine CIK cells.The percentage of CD3＋CD8＋ and CD3＋CD56＋ in CIK cells induced by IL-12 were significantly higher than those in IFN-γ induced CIK（P0.05）,and the percentage of CD3＋CD4＋ and CD4/CD8 were significantly lower in IL-12 induced CIK than those in IFN-γ induced CIK（P0.05）.In addition,the cytotoxic activity of IL-12 induced CIK cells was significantly higher than that of IFN-γ induced CIK（P0.05）.It is concluded that there were higher percentages of both effector cells and CD8＋ specific T cells in CIK cells induced by IL-12,suggesting that IL-12 can take place IFN-γ to prepare CIK cells and the CIK cells will be more effective.