活菌H37Ra与灭活菌H37Rv感染小鼠腹腔巨噬细胞的实验研究

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。

Infection of macrophages with viable strain H37Ra and Heat-Killed H37Rv

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 respe...