Department of Bacteriology, Graduate School of Medicine, University of the Ryukyus, Okinawa, Japan.
Department of Bacterial Pathogenesis, Infection and Host Response, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
Methods Mol Biol. 2020;2134:199-206. doi: 10.1007/978-1-0716-0459-5_18.
Macrophages are phagocytic cells that constitute the primary barrier against pathogens. After phagocytosis a single-membraned vesicle that contains the pathogen is formed. This phagosome undergoes a maturation process to acquire an increasingly antimicrobial environment. Leptospiral uptake by macrophages induces the formation of a Leptospira-containing phagosome (LCP). The kinetics of lysosomal marker recruitment by the LCP is correlated with virulence. This chapter presents a protocol to study the intracellular trafficking of Leptospira spp. within macrophages by fluorescent labeling bacteria and different markers of the phagocytic pathway. We also describe a method to evaluate the bacterial survival within macrophages.
巨噬细胞是吞噬细胞，构成了抵御病原体的主要屏障。吞噬作用后，形成一个含有病原体的单膜囊泡。这个吞噬体经历一个成熟过程，以获得一个越来越具有抗菌环境。巨噬细胞摄取钩端螺旋体诱导形成含有钩端螺旋体的吞噬体（LCP）。溶酶体标记物被 LCP 募集的动力学与毒力相关。本章介绍了一种通过荧光标记细菌和吞噬途径的不同标记物来研究钩端螺旋体属在巨噬细胞内的细胞内运输的方案。我们还描述了一种评估细菌在巨噬细胞内存活的方法。
