IFN-γ对体外培养系统中IL-18诱导的肿瘤特异性CTL杀伤效应的影响  

The effect of IFN-γ on the tumor specific CTLs induced by rhIL-18 in the cell co-culture system

作　　者：方艳秋[1] 齐亚灵[1] 孙绍骞[2] 谭岩[1] 段秀梅[1] 许淑芬[1] 

机构地区：[1]吉林大学第一医院中心实验吉林省卫生厅生物治疗及基因诊断重点实验室,长春130021 [2]吉林大学白求恩医学部临床一系,长春130021

出　　处：《中国免疫学杂志》2007年第12期1078-1082,共5页Chinese Journal of Immunology

基　　金：卫生部临床学科重点科研项目(20013145);吉林省科技厅国际合作项目(20040707-2);吉林省杰出青年资助基金(20050113)项目

摘　　要：目的:应用rhIL-18在体外培养系统(Coculture system in vitro,CCs)中诱导肿瘤特异性细胞毒性T淋巴细胞(Cytotoxic TLymphocyte,CTL),探讨IFN-γ在rhIL-18诱导的肿瘤特异性CTL产生过程及杀伤效应中的作用。方法:采用StemSepTM免疫磁性细胞分离法分离人外周血NK细胞、T细胞及DCs细胞,流式细胞仪分析细胞表型,125I-UdR标记的细胞毒实验检测杀伤活性,ELISA方法检测IFN-γ蛋白产生量,RT-PCR检测IFN-γmRNA表达含量。结果:在肿瘤抗原存在的条件下,IL-18在CCs中能够诱导并促进CTL介导的肿瘤特异性杀伤效应;IL-18能够在肿瘤抗原刺激的CCs中诱导IFN-γmRNA的表达及IFN-γ蛋白的产生,并与IL-18的含量呈剂量依赖关系,在rhIL-18含量为100ng时,培养上清中IFN-γ为4410±210pg/ml。但加入抗IFN-γ抗体对rhIL-18诱导的肿瘤特异性CTL产生过程无明显影响,不能抑制IL-18诱导的这种肿瘤特异性CTL的杀伤效应。结论:IL-18能够在肿瘤抗原刺激的CCs中有效地诱导CTL介导的肿瘤特异性杀伤效应,并诱导IFN-γ的产生,但其诱导肿瘤特异性CTL的产生过程及杀伤效应与IFN-γ无关。Objective:To investigate the effects of IFN-γ on the tumor specific CTLs induced by rhIL-18 in the in vitro cell co-culture system.Methods:The NK cells, T cells and dendritic cells were separated from fresh PBMCs by using the Stem SepTM immunomagnetic beads. Cell phenotypes of the purified cell populations were identified with FCM technique. The cell co-culture system was established as follows: the cell preparations in which definite cell subset had been deleted was co-cultured with mitotic-inactivated tumor cells in the presence of rhIL-18. Production of IFN-γ was measured by ELISA, IFN-γ mRNA was measured RT-PCR. Cytotoxicities of the various effector cell preparations to a series of tumor cell lines were examined by the isotope releasing assay.Results:In the in vitro cell co-culture system with mitotic-inactivated tumor cells in the presence, rhIL-18 alone and dose-dependently induced tumor specific cytotoxicities. Addition of rhIL-18 alone also stimulated the expression and secretion of IFN-γ. It was surprisingly noticed that moAb specific to IFN-γ did not interfere with the induced tumor specific cytotoxicities.Conclusion:IL-18 could effectively induce the tumor specific CTLs in the in vitro cell co-culture system, which may not depend on the modulation of inducible IFN-γ production.

关 键 词：体外培养系统 IL-18 肿瘤杀伤效应 细胞毒性T淋巴细胞 IFN-Γ 

分 类 号：R932.1[医药卫生—生药学] R730.3[医药卫生—药学]