IFN-γ增强人γδT细胞杀伤骨肉瘤细胞的研究

Interferon-γ enhances human γδ T cell-mediated osteosarcoma cell killing in vitro

目的研究干扰素-γ(IFN-γ)增强骨肉瘤患者γδT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γδT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γδT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γδT细胞杀伤OS细胞作用影响较小(P>0.05);但能显著阻断γδT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γδT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 ofγδ 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 cytotoit icity of γδ T ceils in untreated osteosarcoma targets （P〉0.05）, but significantly impaired γδT cell cytotoxicity in IFN-γ pre-treated osteosarcoma targets （P〈0.01）. Conclusion IFN-y can enhance the cytotoxic effect of human γδ T cells from osteosarcorna patients against osteosarcoma cells in vitro