中文摘要：

目的：探讨小鼠腹腔接种结核分枝杆菌活菌H37Ra、灭活菌H37Rv后,诱导巨噬细胞免疫物质的表达差异,探讨活菌H37Ra及灭活菌H37Rv作为新的结核候选疫苗的可行性。方法：分别培养BALB/c小鼠腹腔巨噬细胞,用活菌H37Ra,灭活菌H37Rv感染10小时后用抗酸染色方法,观察各组巨噬细胞吞噬情况并计算吞噬率。将活菌H37Ra、灭活菌H37Rv、生理盐水分别接种BALB/c小鼠腹腔,免疫30天后取小鼠腹腔巨噬细胞,RT-PCR法检测小鼠腹腔巨噬细胞IL-12P40、TNFα-、IFNγ-的基因转录水平;ELISA法检测细胞因子IL-12P40、TNFα-、IFNγ-的表达;Griess法、化学法检测小鼠腹腔巨噬细胞分泌NO、H2O2水平;流式细胞仪检测IFNγ-诱导的CD40L的变化情况。结果：巨噬细胞对活菌H37Ra和灭活菌H37Rv的吞噬率分别为（55.71±8.42）%、（14.82±2.12）%。与灭活菌相比,活菌H37Ra有更强的被吞噬能力（P〈0.01）。活菌H37Ra免疫小鼠腹腔巨噬细胞后能显著诱导巨噬细胞的IL-12P40、TNF及IFNγ-基因的转录;细胞因子IL-12P40、TNFα-、IFNγ-、NO及H2O2高水平分泌;同时活菌H37Ra刺激IFNγ-诱导的巨噬细胞表面CD40L的高表达。结论：活的结核分枝杆菌H37Ra能显著诱导小鼠腹腔巨噬细胞产生更多的保护性免疫物质,有利于免疫应答,有可能作为新的结核候选疫苗,而灭活菌H37Rv不能显著激活小鼠腹腔巨噬细胞分泌保护性免疫物质,不宜作为新的结核候选疫苗。

英文摘要：

Objective：To study the difference in the expression of immune substances which macrophages are induced by inoculating mice with viable M.tuberculosis H37Ra and Heat-Killed M.tuberculosis H37Rv,and to compare the difference between viable and Heat-Killed M.tuberculosis in the induction of immune responses,and thus to learn whether viable or Heat-Killed bacteria could be a possible candidate for new vaccines.Methods：Peritoneal macrophages from BALB/c mouse were cultured with H37Ra and H37Rv for 10 hours respectively,to observe the situation of each group macrophages,and calculate the phagocytic rates of macrophages by the acid dye staining method.BALB/c mice were abdominally immunized with viable H37Ra,Heat-Killed H37Rv and NS,respectively.At 30 days after immunization,the macrophages were isolated.The expression levels of IL-12P40,TNF-α,IFN-γ were determined by RT-PCR and ELISA.The expression levels of NO,H2O2 were determined by Griess and Chemical method;and the changes of CD40L induced IFN-γ were detected by flow cytometry.Results：The phagocytic rates of macrophages were（55.71±8.42）% for viable M.tuberculosis and（14.82±2.12）% for Heat-Killed M.tuberculosis with a significant difference（P0.01）.The mRNA levels of IL-12P40,TNF-α,IFN-γ were higher in viable M.tuberculosis H37Ra than those of in Heat-Killed M.tuberculosis,the cytokine levels of IL-12P40,TNF-α,IFN-γ,NO and H2O2 were higher in Viable M.tuberculosis H37Ra than those of in Heat-Killed M.tuberculosis.At the same time,the expressions of CD40L on macrophages induced by IFN-γ in viable M.tuberculosis stimulated group was higher than that of Heat-Killed M.tuberculosis stimulated group.Conclusion：Viable M.tuberculosis H37Ra can induce the activation of macrophages and the production of more protective immune substances,which play important roles in the host immune response,so it would be a candidate for tuberculosis vaccine.However,Heat-Killed M.tuberculosis H37Rv can＇t induce the activation of macrophages effectively and produc