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目的观察注射用母牛分枝杆菌(微卡)对结核菌感染小鼠细胞免疫功能的影响,并探讨其作用机制。方法C57小鼠经尾静脉注射结核分枝杆菌H37RV,建立结核菌感染小鼠模型。将模型小鼠随机分为模型组、微卡高、中、低3个剂量组和异烟肼组,同时设正常对照组。感染8周后处死小鼠,取肺、脾、肝,计算重量指数和病变总指数,测定肺脏活菌数量,并观察主要脏器的病理变化;流式细胞术测定小鼠脾脏中CD3+T、CD4+T、NK1.1+T、NK、γδT细胞的比例以及CD4+T细胞胞内IFNγ和IL-4的表达。结果微卡各剂量组能明显减轻感染小鼠的发病状况,其中微卡中剂量组的各脏器病变总指数、脾脏和肺脏重量指数、肺脏结核菌活菌数均显著低于模型组;微卡各剂量组小鼠的肺部病变主要以增殖性结节为主,而模型组则以坏死性结节和增殖性结节为主;微卡各剂量组小鼠的CD3+T、CD4+T细胞比例均明显升高,CD4+IFNγ+辅助性T细胞比例与模型组比较,差异无统计学意义;微卡低、中剂量组CD4+IL-4+辅助性T细胞比例明显降低,同时能显著提高小鼠天然免疫系统γδT和NK细胞的比例。结论微卡可提高结核菌感染小鼠天然和获得性细胞免疫水平,对结核菌感染小鼠具有较好的免疫干预作用。
Objective To observe the effects of Mycobacterium vaccae injection (Mika) on cellular immune function in mice infected with Mycobacterium tuberculosis and to explore its mechanism. Methods C57 mice were injected with Mycobacterium tuberculosis H37RV through tail vein to establish a mouse model of Mycobacterium tuberculosis infection. The model mice were randomly divided into model group, micro-card high, medium and low dose groups and isoniazid group, while the normal control group. The mice were sacrificed 8 weeks after infection, lungs, spleen and liver were taken out, the weight index and total lesion index were calculated, the number of viable cells in the lungs was measured, and the pathological changes of the major organs were observed. Flow cytometry was used to detect CD3 + T , CD4 + T, NK1.1 + T, NK, γδ T cells and intracellular IFNγ and IL-4 expression of CD4 + T cells. Results Each dose of microcapsules could significantly reduce the incidence of infected mice. The total lesion index, weight index of spleen and lungs, and viable count of pulmonary tuberculosis in the Mika medium dose group were significantly lower than those in the model group. Cardiomyopathy mice were predominantly of proliferative nodules in lung lesions and necrotic nodules and proliferative nodules in the model group. CD3 + T and CD4 + The percentage of CD4 + IL-4 + T-cells in CD4 + IFN-γ + T helper cells was significantly lower than that in the untreated group Can significantly increase the ratio of γδT and NK cells in mice’s natural immune system. Conclusion Micro-card can enhance the natural and acquired cellular immunity of mice infected with Mycobacterium tuberculosis, and has good immunological intervention on mice infected with TB.