【摘要】 目的探索IL-15对CIK细胞NKG2D表达及对肿瘤细胞体外杀伤活性的影响。方法以常规培养的CIK细胞(常规组)为对照,流式细胞术(FCM)检测常规培养体系中加入IL-15后CIK细胞(IL-15组)的NKG2D表达情况;MTT法检测IL-15组及封闭NKG2D受体的CIK细胞对MDA-MB-231、SKBr-3等癌细胞的杀伤活性。结果①培养后的各个时段,IL-15组CIK细胞中T淋巴细胞亚群、CD3+CD56+细胞及总CIK细胞群的NKG2D表达水平高于常规组,两组的差异有统计学意义(P<0.05)。②培养14、21d的IL-15组CIK细胞对MDA-MB-231和SKBr-3的杀伤活性明显高于常规组,差异有统计学意义(P<0.05)。③NKG2D单抗可以下调CIK细胞对MDA-MB-231细胞的杀伤效应,而对SKBr-3细胞的杀伤没有明显影响。结论 IL-15可以增加CIK细胞NKG2D的表达水平并提高对肿瘤细胞的杀伤活性,NKG2D途径是CIK细胞杀伤肿瘤细胞的重要机制之一。更多还原

【Abstract】 Objective This aim of this study was to investigate the effect of interleukin-15(IL-15) on the NKG2D expression of cytokine induced killers(CIKs) and the cytotoxicity of CIKs against breast cancer cells in vitro.Methods Mononuclear cells were isolated from peripheral blood of 15 healthy volunteers by Ficoll density gradient centrifugation,and were induced to differentiate into CIKs in the presence of various cytokines with or without IL-15.Cells were harvested each week for 4 weeks starting with day 0,and NKG2D protein levels were determined using flow cytometry.The cytotoxicity of CIKs against breast cancer cells MDA-MB-231 and SKBr-3 was assessed using MTT assay.Results T lymphocyte subpopulation,CD3 + CD5 6 + CIKs and the total CIKs from the IL-15-treated group showed significantly higher levels of NKG2D compared to the counterparts from the IL-15-untreated group(P<0.05) .IL-15-treated CIKs demonstrated a stronger cytotoxicity against MDA-MB-231 and SKBr-3 than those without the IL-15 treatment(P<0.05) .Most interestingly,the cytotoxic effects of CIKs on MDA-MB-231 cells were considerably reduced by the pretreatment with anti-NKG2D monoclonal antibody.However,the treatment with the same antibody had little influence on the cytotoxicity of CIKs against SKBr-3 cells.Conclusion IL-15 enhances the expression of NKG2D on the surface of CIKs and increases the cytotoxicity of CIKs against breast cancer cells.NKG2D plays an important role in mediating the cytotoxicity of CIKs on specific types of tumor cells.更多还原