摘要
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12~24h,诱导作用在16~18h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 TB10.4and RAW264.7cells,recombinant TB10.4was expressed and purified,and the cell activities were detected by IFN-γrelease assays and Western blot.RAW264.7cells 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 rTB10.4on the expression and distribution of TLR2 of RAW264.7cells were detected and analyzed by Image-Stream fluorescence imaging.The results showed that the rTB10.4has strong T cells and B cell response activities.The interaction of RAW264.7with rTB10.4was dose-dependent,and the onset time range was from 12 hto 24h,and the maximum effect was observed from16 hto 18h.The results of Image-Stream fluorescence imaging showed that the stimulation of rTB10.4could significantly enhance the expression of TLR2 on the cell membrane of RAW264.7cells.In a word,this study provided a scientific basis for further exploring the TB10.4protein function and its role on the diagnosis of bovine tuberculosis.
出处
《华中农业大学学报》
CAS
CSCD
北大核心
2015年第1期83-90,共8页
Journal of Huazhong Agricultural University
基金
国家自然科学基金项目(31302130)
"中国农业科学院科技创新工程"兽医公共卫生安全与管理创新团队(ASTIP-IAS-11)
国家"863"高技术研究与发展专项(2012AA101302)
