| High-throughput Microscopic Analysis of Salmonella Invasion of Host Cells
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Author:
Date:
2018-09-20
[Abstract] Salmonella is a Gram-negative bacterium causing a gastro-enteric disease called salmonellosis. During the first phase of infection, Salmonella uses its flagella to swim near the surface of the epithelial cells and to target specific site of infection. In order to study the selection criteria that determine which host cells are targeted by the pathogen, and to analyze the relation between infecting Salmonella (i.e., cooperation or competition), we have established a high-throughput microscopic assay of HeLa cells sequentially infected with fluorescent bacteria. Using an automated pipeline of image analysis, we quantitatively characterized a multitude of parameters of infected and non-infected cells. Based on this, we established a predictive model that ...
[摘要] 沙门氏菌是革兰氏阴性细菌，引起称为沙门氏菌病的胃肠疾病。在感染的第一阶段，沙门氏菌使用其鞭毛在上皮细胞表面附近游泳并靶向特定的感染部位。为了研究确定哪种宿主细胞被病原体靶向的选择标准，并分析感染沙门氏菌（ ie ，合作或竞争）之间的关系，我们有建立了对荧光细菌依次感染的HeLa细胞的高通量显微镜检测。使用自动化图像分析管道，我们定量表征了感染和未感染细胞的众多参数。基于此，我们建立了一个预测模型，使我们能够识别宿主细胞易受感染的参数。我们发现宿主细胞易损性有两个来源：病原体诱导的细胞易感性从沙门氏菌摄取中出现并持续存在于感染过程的后期阶段;以及与细胞固有属性相关的宿主细胞固有的脆弱性，例如局部细胞拥挤和胆固醇含量。我们的方法基于形态学或分子宿主细胞参数预测单层上皮细胞中沙门氏菌感染的概率。在这里，我们提供了工作流程的详细描述，包括基于计算机的分析管道。我们的方法有可能应用于研究宿主 - 病原体相互作用的其他组合。
【背景】鼠伤寒沙门氏菌血清型鼠伤寒沙门氏菌通过摄入受污染的食物或水感染宿主，引起沙门氏菌病。一旦细菌到达肠道的远端回肠，它们就会侵入广泛的宿主细胞，包括肠上皮细胞（Watson和Holden，2010）。在宿主细胞入侵的第一阶段，沙门氏菌选择其目标，使用其鞭毛游泳并扫描上皮表面（Misselwitz et ...
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| Design of a Transcription-based Secretion Activity Reporter (TSAR) for the Type III Secretion Apparatus of Shigella flexneri and Uses Thereof
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Author:
Date:
2014-10-20
[Abstract] Many gram-negative bacterial pathogens, including Shigella flexneri, are able to translocate bacterial proteins, dubbed effectors, across the host cell plasma membrane into the host cell cytosol using a syringe-like structure, the type three secretion apparatus (T3SA). While some bacteria use their T3SA to modulate their phagosomal environment (Salmonella spp.), establish pedestal structure to form microcolonies on the plasma membrane (Enteropathogenic Escherichi coli) or lyse their entry vacuole (Shigella spp.), they all have in common a tightly regulated activity of their T3SA. However, the tracking of the activity of the T3SA in infected cells and tissue has been difficult to perform. Using the property of MxiE-dependent promoters that are ...
[摘要] 包括灵芝氏菌在内的许多革兰氏阴性细菌病原体能够使用注射器样结构将类型三分泌物穿过宿主细胞质膜转运细菌蛋白质（配体效应子）到宿主细胞胞质溶胶中装置（T3SA）。虽然一些细菌使用它们的T3SA调节它们的吞噬体环境（沙门氏菌 spp 。），建立基质结构以在质膜上形成微集落（Enteropathogenic < em="">）或溶解它们的进入泡（志贺氏菌属。），它们都具有共同的严格调节的他们的T3SA的活性。然而，T3SA在感染的细胞和组织中的活性的跟踪一直难以进行。使用在T3SA是活性时上调的MxiE依赖性启动子的性质，我们最近设计了基于转录的分泌活性报道分子（TSAR），其允许以下的S的活性。使用快速成熟的GFP内在荧光，在组织培养细胞中实时地和体内实时分析灵敏度。在这里我们描述TSAR的设计及其应用于固定和活体样品的显微镜和流式细胞仪在结肠上皮细胞模型使用TC7组织培养细胞。
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