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目的:探讨重组耻垢分枝杆菌Ag85B-ESAT6-rMs对小鼠巨噬细胞功能的影响。方法:将耻垢分枝杆菌(Ms)与重组耻垢分枝杆菌Ag85B-ESAT6-rMs分别感染小鼠巨噬细胞系RAW264.7,通过抗酸染色和细胞内细菌菌落形成单位计数评价Ms和Ag85B-ESAT6-rMs被巨噬细胞吞噬和消化能力;采用流式细胞术检测巨噬细胞凋亡情况;分别采用实时定量PCR技术和ELISA检测细胞因子TNF-α的表达和分泌水平。结果:小鼠巨噬细胞实验证实,Ag85B-ESAT6-rMs比Ms具有更强的感染和促进巨噬细胞的吞噬能力,Ag85B-ESAT6-rMs可增加巨噬细胞表达和分泌TNF-α的水平,并促进受感染巨噬细胞的凋亡。结论:Ag85B-ESAT6-rMs可以促进巨噬细胞的吞噬消化功能,将有助于增强巨噬细胞对Ms抗原的递呈和机体抗Ms感染的特异性免疫。
Objective: To investigate the effect of recombinant Mycobacterium smegmatis Ag85B-ESAT6-rMs on the function of mouse macrophages. Methods: Mycobacterium smegmatis (Ms) and recombinant Mycobacterium smegmatis Ag85B-ESAT6-rMs were transfected into mouse macrophage cell line RAW264.7, respectively, and MS and MSCs were evaluated by acid-fast staining and intracellular bacterial colony forming units Ag85B-ESAT6-rMs were phagocytosed and digested by macrophages. The apoptosis of macrophages was detected by flow cytometry. The expression and secretion of cytokine TNF-α were detected by real-time PCR and ELISA. Results: Ag85B-ESAT6-rMs showed stronger infection and promoted macrophage phagocytosis than M85B-ESAT6-rMs. Ag85B-ESAT6-rMs increased the level of TNF-α expressed and secreted by macrophages, And promote the apoptosis of infected macrophages. Conclusion: Ag85B-ESAT6-rMs can promote macrophage phagocytosis and digestion, which will help to enhance the specific immunity of macrophages to Ms antigen presentation and anti-Ms infection.