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Plaque Assay to Determine Invasion and Intercellular Dissemination of Shigella flexneri in TC7 Human Intestinal Epithelial Cells
Umeå universitet, Medicinska fakulteten, Molekylär Infektionsmedicin, Sverige (MIMS). Umeå universitet, Medicinska fakulteten, Umeå Centre for Microbial Research (UCMR). Umeå universitet, Medicinska fakulteten, Institutionen för molekylärbiologi (Medicinska fakulteten).
Umeå universitet, Medicinska fakulteten, Molekylär Infektionsmedicin, Sverige (MIMS). Umeå universitet, Medicinska fakulteten, Umeå Centre for Microbial Research (UCMR). Umeå universitet, Medicinska fakulteten, Institutionen för molekylärbiologi (Medicinska fakulteten).ORCID-id: 0000-0002-9915-002x
2019 (engelsk)Inngår i: BIO-PROTOCOL, ISSN 2331-8325, Vol. 9, nr 13, artikkel-id UNSP e3293Artikkel i tidsskrift (Fagfellevurdert) 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.

sted, utgiver, år, opplag, sider
BIO-PROTOCOL , 2019. Vol. 9, nr 13, artikkel-id UNSP e3293
Emneord [en]
Intracellular bacteria, Intestinal epithelial cells, Giemsa stain, Shigella, Invasion, Dissemination
HSV kategori
Identifikatorer
URN: urn:nbn:se:umu:diva-162002DOI: 10.21769/BioProtoc.3293ISI: 000476647800010OAI: oai:DiVA.org:umu-162002DiVA, id: diva2:1342250
Tilgjengelig fra: 2019-08-13 Laget: 2019-08-13 Sist oppdatert: 2019-08-13bibliografisk kontrollert

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