Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, P.O. Box 56, FI-00014, University of Helsinki, Finland.
Drug Research Program, Division of Pharmaceutical Biosciences, Faculty of Pharmacy, P.O. Box 56, FI-00014, University of Helsinki, Finland.
J Microbiol Methods. 2021 May;184:106201. doi: 10.1016/j.mimet.2021.106201. Epub 2021 Mar 10.
Enteropathogenic E. coli (EPEC) causes intestinal infections leading to severe diarrhea. EPEC attaches to the host cell causing lesions to the intestinal epithelium coupled with the effacement of microvilli. In the process, actin accumulates into a pedestal-like structure under bacterial microcolonies. We designed an automated fluorescence microscopy-based screening method for discovering compounds capable of inhibiting EPEC adhesion and virulence using aurodox, a type three secretion system (T3SS) inhibitor, as a positive control. The screening assay employs an EPEC strain (2348/69) expressing a fluorescent protein and actin staining for monitoring the bacteria and their pedestals respectively, analyzing these with a custom image analysis pipeline. The assay allows for the discovery of compounds capable of preventing the formation of pathogenic actin rearrangements. These compounds may be interfering with virulence-related molecular pathways relevant for developing antivirulence leads.
肠致病性大肠杆菌（EPEC）可引起严重腹泻的肠道感染。EPEC 附着在宿主细胞上，导致肠上皮损伤，微绒毛消失。在此过程中，肌动蛋白在细菌微菌落下积累成墩状结构。我们设计了一种基于自动化荧光显微镜的筛选方法，使用三型分泌系统（T3SS）抑制剂aurodox 作为阳性对照，来发现能够抑制 EPEC 黏附和毒力的化合物。该筛选检测方法采用表达荧光蛋白和肌动蛋白染色的 EPEC 菌株（2348/69），分别用于监测细菌及其墩的形成，使用自定义的图像分析管道进行分析。该检测方法可发现能够防止致病性肌动蛋白重排形成的化合物。这些化合物可能干扰与开发抗毒力先导物相关的毒力相关分子途径。
