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Plaque Assay to Determine Invasion and Intercellular Dissemination of Shigella flexneri in TC7 Human Intestinal Epithelial Cells
Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS). Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR). Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine).
Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS). Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR). Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine).ORCID iD: 0000-0002-9915-002x
2019 (English)In: BIO-PROTOCOL, ISSN 2331-8325, Vol. 9, no 13, article id UNSP e3293Article in journal (Refereed) Published
Abstract [en]

Shigella flexneri invades the epithelial cells lining the gut lumen and replicates intracellularly. The specialized Type III Secretion System (T3SS) and its effector proteins, encoded on a large virulence plasmid, assist the bacterium to gain access to the cytosol. Thereafter Shigella disseminates to neighboring cells in an epithelial layer without further extracellular steps. Host cell lysis occurs when these bacteria have extensively replicated in the target cell cytosol. Here we describe a simple method to qualitatively as well as quantitatively study the capacity of Shigella to invade and disseminate within an epithelium by assessing the number and size of plaques representing the dead cells in a monolayer of TC7 cells. This classical protocol follows a simple approach of infecting the monolayers of epithelial cell lines with Shigella and visualizing the dead cells as plaques formed against a stained background.

Place, publisher, year, edition, pages
BIO-PROTOCOL , 2019. Vol. 9, no 13, article id UNSP e3293
Keywords [en]
Intracellular bacteria, Intestinal epithelial cells, Giemsa stain, Shigella, Invasion, Dissemination
National Category
Immunology in the medical area
Identifiers
URN: urn:nbn:se:umu:diva-162002DOI: 10.21769/BioProtoc.3293ISI: 000476647800010OAI: oai:DiVA.org:umu-162002DiVA, id: diva2:1342250
Available from: 2019-08-13 Created: 2019-08-13 Last updated: 2019-08-13Bibliographically approved

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Sharma, AtinPuhar, Andrea

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Molecular Infection Medicine Sweden (MIMS)Umeå Centre for Microbial Research (UCMR)Department of Molecular Biology (Faculty of Medicine)
Immunology in the medical area

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