IFN-γ增强人γδ T细胞杀伤骨肉瘤细胞的研究

摘要

目的 研究干扰素-γ(IFN-γ)增强骨肉瘤患者γδ T细胞对骨肉瘤杀伤作用的影响.方法 以唑来磷酸法扩增骨肉瘤患者外周血γδT细胞.分别使用实时荧光定量聚合酶链反应和流式细胞术检测IFN-γ处理前后骨肉瘤细胞Fas的表达情况；LDH法检测骨肉瘤患者γδ T细胞对骨肉瘤细胞的杀伤活性.结果 IFN-γ对骨肉瘤细胞Fas的表达明显提高(P＜0.01),骨肉瘤患者γδ T细胞对骨肉瘤细胞的杀伤作用显著增强(P＜0.01).抗人FasL抗体对γδ T细胞杀伤OS细胞作用影响较小(P＞0.05)；但能显著阻断γδ T细胞杀伤IFN-γ预处理OS细胞的作用,差异均有统计学意义(P＜0.01).结论 IFN-γ可增强骨肉瘤患者γδ T细胞的抗骨肉瘤作用.%Objective To investigate the cytotoxic effect of γδ T cells from osteosarcoma patients against interferon-γ (IFN-γ)-treated osteosarcoma cells in vitro. Methods Human γδ T cells were amplified by zoledronate from peripheral blood cells of osteosarcoma patients. The expression of Fas on the osteosarcoma cells were measured by flow cytometry and quantitative real-time PCR analysis before and after IFN-γ treatment. The cytoxicity of γδ T cells against osteosarcoma cells was evaluated with LDH assay. Results IFN-γ significantly enhanced the susceptibility of the osteosarcoma cell lines HOS and U2OS to the cycotoxicity of γδ T cells from osteosarcoma patients (P<0.01). IFN-γ obviously up-regulated the expression of Fas in HOS and U2OS cells (P<0.01). Anti-FasL mAb failed to inhibit the cytotoxicity of γδ T cells in untreated osteosarcoma targets (P>0.05), but significantly impaired γδ T cell cytotoxicity in IFN-γ pre-treated osteosarcoma targets (P<0.01). Conclusion IFN-γ can enhance the cytotoxic effect of human γδ T cells from osteosarcoma patients against osteosarcoma cells in vitro