Centre for Innate Immunity and Infectious Diseases, Hudson Institute of Medical Research, Clayton, VIC, Australia.
Department of Molecular and Translational Science, Faculty of Medicine, Nursing and Health Sciences, Monash University, Clayton, VIC, Australia.
Methods Mol Biol. 2022;2523:23-41. doi: 10.1007/978-1-0716-2449-4_3.
Legionella pneumophila is an intracellular bacterial pathogen that uses a type IV secretion system (T4SS), termed Dot/Icm, to secrete more than 330 virulence effector proteins into the infected host cell. Many Dot/Icm effectors are involved in biogenesis of the Legionella-containing vacuole (LCV), which allows intracellular bacterial replication in environmental amoebae and alveolar macrophages. Through their activity, some effectors trigger the mammalian host immune response in a phenomenon termed effector-triggered immunity (ETI). Here, we describe a protocol to create and use L. pneumophila genome deletion mutants to identify effector(s) that alter pro-inflammatory cytokine production and bacterial clearance in the lungs of mice.
嗜肺军团菌是一种细胞内细菌病原体，它利用一种称为 Dot/Icm 的 IV 型分泌系统将超过 330 种毒力效应蛋白分泌到感染的宿主细胞中。许多 Dot/Icm 效应蛋白参与包含军团菌的空泡（LCV）的生物发生，这使得细菌能够在环境中的变形虫和肺泡巨噬细胞中进行复制。通过它们的活性，一些效应蛋白在一种称为效应蛋白触发免疫（ETI）的现象中引发哺乳动物宿主的免疫反应。在这里，我们描述了一种创建和使用嗜肺军团菌基因组缺失突变体的方案，以鉴定改变小鼠肺部促炎细胞因子产生和细菌清除的效应蛋白。
