Author:
Date:
2012-10-20
[Abstract] Escherichia coli (E. coli) produces disseminated infections of the urinary tract, blood, and central nervous system where it encounters professional phagocytes such as macrophages, which utilize reactive nitrogen intermediates (RNI) to arrest bacteria. In vitro, extraintestinal pathogenic E. coli (ExPEC) can survive within bone marrow-derived macrophages for greater than 24 h post-infection within a LAMP1+ vesicular compartment, and ExPEC strains, in particular, are better adapted to intracellular macrophage survival than commensal strains (Bokil et al., 2011). This protocol details an intracellular murine macrophage-like cell infection, including modulation of the host nitrosative stress response, to model this host-pathogen interaction in ...
[摘要] 大肠杆菌(大肠杆菌)产生尿道,血液和中枢神经系统的扩散感染,其中它遇到专业吞噬细胞,例如巨噬细胞,其利用反应性氮中间体RNI)以阻止细菌。 体外,肠外致病性。大肠杆菌(ExPEC)可以在LAMP1 +囊泡区室内在感染后在骨髓来源的巨噬细胞中存活超过24小时,并且ExPEC菌株特别地比共生菌株更适合细胞内巨噬细胞存活(Bokil& em>等人,2011)。该协议详细描述了细胞内鼠类巨噬细胞样细胞感染,包括调节宿主亚硝基化应激反应,在体外模拟这种宿主 - 病原体相互作用。为了实现这一点,将RAW 264.7鼠巨噬细胞样细胞与L-精氨酸,NO前体或IFNγ预温育以产生高一氧化氮(NO)生理状态或L-NAME,诱导型NO合酶(iNOS )特异性抑制剂,产生低NO生理状态。该方案已经成功地用于评估新型ExPEC调节剂对巨噬细胞感染期间的细胞内存活和亚硝基化应激反应的贡献(Bateman和Seed,2012),但是可以适用于多种E。大肠杆菌菌株或同基因缺失。
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