牛分枝杆菌重组蛋白TB10.4对RAW264.7细胞TLR2表达和分布的影响
Mycobacterium bovis antigen protein rTB10.4 and its effects on expression and distribution of TLR2 in RAW264.7 cells

为探讨牛分枝杆菌TB10.4蛋白与RAW264.7细胞的相互作用关系，表达纯化了rTB10.4，并采用 IFN-γ释放试验和Western blot检测其细胞活性。使用不同剂量rTB10.4与RAW264.7细胞共孵育后，采用实时无标记动态细胞分析技术检测rTB10.4对RAW264.7细胞生长影响的最佳时间点和最佳剂量。在此基础上，采用量化流式细胞仪分析重组蛋白rTB10.4对RAW264.7细胞TLR2表达的影响。结果表明，获得的rTB10.4具有较强的T细胞和B细胞活性，rTB10.4对RAW264.7细胞的作用呈剂量依赖性,作用起效时间为12~24 h，诱导作用在16~18 h达到最大效应值。量化流式分析仪分析结果表明，rTB10.4刺激可显著增强RAW264.7细胞的细胞膜上TLR2的表达。
To preliminarily explore the interaction between Mycobacterium bovis TB10.4 and RAW264.7 cells,recombinant TB10.4 was expressed and purified,and the cell activities were detected by IFN-γ release assays and Western blot.RAW264.7 cells were incubated with different doses of rTB10.4,then the best exposure condition was analysed by the RT-CES system.On this basis,the effects of rTB10.4 on the expression and distribution of TLR2 of RAW264.7 cells were detected and analyzed by Image-Stream fluorescence imaging.The results showed that the rTB10.4 has strong T cells and B cell response activities.The interaction of RAW264.7 with rTB10.4 was dose-dependent,and the onset time range was from 12 h to 24 h,and the maximum effect was observed from16 h to 18 h.The results of Image-Stream fluorescence imaging showed that the stimulation of rTB10.4 could significantly enhance the expression of TLR2 on the cell membrane of RAW264.7 cells.In a word,this study provided a scientific basis for further exploring the TB10.4 protein function and its role on the diagnosis of bovine tuberculosis．