Proteomic analysis of macrophage in response to Edwardsiella tarda-infection
Proteomic analysis of macrophage in response to Edwardsiella tarda-infection
复制标题
巨噬细胞响应迟缓爱德华氏菌感染的蛋白质组学分析
DOI：
10.1016/j.micpath.2017.08.028
复制
发表时间：
2017-10-01
影响因子：
3.8
通讯作者：
Gao, Yingli
中科院分区：
文献类型：
--
作者：
Qin, Lei;Li, Fuhou;Gao, Yingli
迟发爱德华菌是一种重要的兼性细胞内病原体，感染范围广泛，从鱼类到人类。这种细菌可以在巨噬细胞中存活和复制，作为一种逃避宿主防御的机制。迟缓-巨噬细胞相互作用在决定爱德华菌病的结果中至关重要。为了充分阐明迟发芽孢杆菌的发病机制，采用二维凝胶电泳（2-DE）和液相色谱-串联质谱（LC-MS/MS）技术，分析了不同时间点RAW264.7细胞对迟发芽孢杆菌感染反应的差异蛋白质组。成功鉴定出26个改变蛋白（上调18个，下调8个），主要参与吞噬体的形成、巨噬细胞的杀微生物活性和抗巨噬细胞凋亡。采用实时荧光定量PCR （qPCR）技术对差异表达蛋白的6个对应基因进行定量分析，检测其转录谱。Western blot分析进一步证实了5个蛋白在蛋白质组学谱中的差异表达。基于这些发现，我们假设这些差异表达的蛋白可能在决定迟发E.感染的过程中起关键作用。结果表明，迟缓芽孢杆菌可以通过调节吞噬体的生物发生、对巨噬细胞杀微生物剂的耐药性和抗巨噬细胞凋亡等途径来实现细胞内的生存。因此，这项工作有效地提供了有用的和新的蛋白质相关信息，以进一步了解迟发E.感染的潜在发病机制。(C) 2017 Elsevier Ltd.版权所有。
Edwardsiella tarda is an important facultative intracellular pathogen infecting a wide range of host from fish to humans. This bacterium could survive and replicate in macrophages as an escape mechanism from the host defense. E. tarda-macrophage interaction is vital in determining the outcome of edwardsiellasis. To fully elucidate the pathogenesis of E. tarda, the differential proteomes of RAW264.7 cells in response to E. tarda-infection, were analyzed at different time points with two-dimensional gel electrophoresis (2-DE) followed by liquid-chromatography-tandem mass spectrometry (LC-MS/MS) identification. 26 altered proteins (18 up-regulated and 8 down-regulated proteins) were successfully identified, which are mainly involved in formation of phagosomes, macrophage microbicidal activity and anti-apoptosis of macrophage. Moreover, 6 corresponding genes of the differentially expressed proteins were quantified by quantitative real-time PCR (qPCR) to examine the transcriptional profiles. Western blot analysis further confirmed the differential expression of 5 proteins in the proteomic profiles. Based on these findings, we hypothesize that these differentially expressed proteins likely play a pivotal role in determining the course of E. tarda-infection. The result suggested that E. tarda could develop some strategies to achieve a successful intracellular lifestyle, including modulation of phagosome biogenesis, resistance to macrophage microbicidal agent and anti-apoptosis of macrophages. Thus, this work effectively provides useful and novel protein-related information to further understand the underlying pathogenesis of E. tarda-infection. (C) 2017 Elsevier Ltd. All rights reserved.