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目的:研究IFN-γ诱导上调表达的单核细胞MHC-I类链相关分子(MICs)分子对NK细胞的活化作用。方法:采用密度梯离离心法分离人外周血单个核细胞(PBMC),以免疫磁珠法从PBMC中特异性分选单核细胞及NK细胞,以细胞因子IFN-γ、TNF-α刺激单核细胞后,再将单核细胞与NK细胞共培养,以流式细胞术(FCM)检测NK细胞表面CD69分子及胞内IFN-γ表达,以51Cr释放试验检测NK细胞对K562细胞的杀伤效应。结果:IFN-γ上调人单核细胞表面MICs表达;IFN-γ刺激的单核细胞能促进异体NK细胞CD69及胞内IFN-γ表达,能增强NK细胞对K562细胞的杀伤效应;单核细胞的这种效应至少部分依赖于IFN-γ上调的MICs分子,因为用抗MIC抗体封闭或用细胞小室阻断细胞接触,可以明显地抑制NK细胞的活化。结论:IFN-γ上调人单核细胞表面MICs分子介导了NK细胞的活化。
AIM: To investigate the effects of IFN-γ-induced activation of NK cells on MHC-I chain-related molecules (MICs) in monocytes. Methods: Human peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation. Mononuclear cells and NK cells were sorted from PBMC by immunomagnetic beads method. Monocytes and NK cells were stimulated with IFN-γ and TNF-α After mononuclear cells and NK cells were co-cultured, the expression of CD69 and IFN-γ on NK cells was detected by flow cytometry (FCM). The killing effect of NK cells on K562 cells was detected by 51Cr release assay . Results: IFN-γ up-regulated the expression of MICs on human monocytes. IFN-γ-stimulated monocytes promoted the expression of CD69 and IFN-γ in allogeneic NK cells, and enhanced the killing effect of NK cells on K562 cells. Monocytes Of this effect depends, at least in part, on the upregulation of IFN-γ in MICs molecules, as activation of NK cells can be significantly inhibited by blocking with anti-MIC antibodies or by blocking cell contact with the cell compartment. Conclusion: IFN-γ upregulates human monocyte surface MICs molecules and mediates NK cell activation.