【摘要】 目的:研究Toll样配体(R-848)与IL-12对人NK细胞IFN-γ产生的作用和细胞亚群分析。方法:分离人外周血PBMC和纯化的NK细胞,分别与R-848、IL-12或R-848和IL-12共同培养。利用ELISA法检测培养上清中IFN-γ的水平,再利用流式检测并分析产生IFN-γ的NK细胞亚群。结果:正常人PBMC分别与不同浓度的Toll样配体R-848、LPS、CpG培养后,均以剂量依赖的方式诱导IFN-γ的产生,但以R-848的效果最佳。细胞亚群分析的结果表明,R-848对CD4+T和CD8+T细胞IFN-γ的表达无明显作用,但显著地促进CD56+细胞表达IFN-γ。同样地,在IL-12刺激之下,CD56bright和CD56dimNK细胞表达IFN-γ。当R-848和IL-12与PBMC和纯化NK细胞孵育后,对CD56bright和CD56dimNK细胞IFN-γ的表达具有协同作用。结论:Toll样配体与NK细胞Toll样受体结合后,促进CD56brightNK细胞亚群IFN-γ的产生,而且Toll样配体与IL-12具有协同作用,提示Toll样受体与细胞因子在调控NK细胞的生物活性中发挥着十分重要的作用。更多还原

【Abstract】 AIM:To investigate the effect of the TLR ligand(R-848) and IL-12 on the production of IFN-γ by human NK cell subsets.METHODS:PBMC or purified NK cells were isolated from normal human peripheral blood and cultured with R-848,IL-12,R-848+ IL-12.The level of IFN-γ in the culture supernatants was measured by ELISA.The cell subsets which produced IFN-γ were detected and analyzed by flow cytometry(FCM).RESULTS:PBMC cultured with different concentrations of TLR ligands(R-848,LPS and CpG) could induce IFN-γ production in a dose dependent manner.Among these three TLR ligands,R-848 was the best one in induction of IFN-γ production by PBMC.FCM analysis indicated that R-848 could induce IFN-γ expression by CD56+ cells,but not CD4+ or CD8+T cells.Similarly,IL-12 could stimulate NK cells to produce IFN-γ.In addition,R-848 and IL-12 had synergistic effect on the production of IFN-γ by PBMC and purified NK cells including CD56bright and CD56dim subsets.CONCLUSION:Engagement of TLR7/8 by R-848 stimulation with IL-12 could induce IFN-γ production by CD56bright NK cells which indicated that TLRs and cytokines played an important role in the regulation of biologic function of NK cells.更多还原