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目的评估携带小鼠胞内病原体抗性基因1(intracellular pathogen resistance1,Ipr1)质粒(pBOGI)的重组卡介苗(BCG)对结核分枝杆菌(Mtb)感染的治疗效果。方法BALB/c小鼠30只,随机分为3组:分别滴鼻给予磷酸盐缓冲液(PBS)、BCG和重组BCG,2周后用人型Mtb H37Rv标准株感染所有小鼠;感染后分别用PBS、BCG和重组BCG治疗7次;末次治疗后处死小鼠,检测肺脾荷菌量,肺脾脏器指数,血清IFN-γ、IL-10及TNF-α分泌水平和肺脾组织中Ipr1的表达,同时观察小鼠肺脾组织病理改变情况。结果重组BCG组肺脾荷菌量比PBS组和BCG组显著降低(P<0.01);重组BCG组肺脾脏器指数比PBS组和BCG组显著降低(P<0.05);重组BCG组血清IFN-γ分泌水平比PBS组和BCG组显著升高(P<0.01)。用免疫组化检测到小鼠肺脾组织中有Ipr1表达。PBS组肺组织病理改变以渗出为主,病变广泛;重组BCG组肺部病变最轻,肺泡结构基本正常;BCG组病变范围局限,病变程度界于PBS组与重组BCG组之间;各组间比较脾脏病理改变不明显。结论携带小鼠Ipr1基因的重组BCG对结核分枝杆菌感染有一定的治疗作用。
Objective To evaluate the therapeutic effect of recombinant bacille Calmette-Guérin (BCG) harboring mouse intracellular pathogen resistance 1 (pprO) plasmid (pBOGI) on M. tuberculosis (Mtb) infection. Methods Thirty BALB / c mice were randomly divided into three groups: PBS, BCG and recombinant BCG were intranasally administrated, and all mice were infected with the standard Mtb H37Rv two weeks later. After infection, PBS, BCG and recombinant BCG for 7 times. After the last treatment, mice were sacrificed to detect the spleen and spleen load, lung and spleen index, serum levels of IFN-γ, IL-10 and TNF-α and Ipr1 in lung and spleen Expression, at the same time observe the pathological changes of lung and spleen in mice. Results Compared with PBS group and BCG group, the amount of spleen and spleen load in recombinant BCG group was significantly lower than that in PBS group and BCG group (P <0.01); the index of lung and spleen in BCG group was significantly lower than that in PBS group and BCG group (P <0.05) γ secretion was significantly higher than PBS group and BCG group (P <0.01). Ipr1 expression was detected in lung and spleen of mice by immunohistochemistry. In PBS group, the pathological changes of lung tissue were mainly exudation with extensive lesions. The lung lesions in livers of BCG group were the lightest and the alveolar structures were basically normal. The range of lesion in BCG group was limited between PBS group and recombinant BCG group. Pathological changes between the spleen is not obvious. Conclusion Recombinant BCG carrying mouse Ipr1 gene has certain therapeutic effect on Mycobacterium tuberculosis infection.