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．