Determining Shiga Toxin-Producing Escherichia coli Interactions with Human Intestinal Epithelium in a Microaerobic Vertical Diffusion Chamber

Conor J McGrath; Stephanie Schüller

doi:10.1007/978-1-0716-1339-9_12

Determining Shiga Toxin-Producing Escherichia coli Interactions with Human Intestinal Epithelium in a Microaerobic Vertical Diffusion Chamber

Methods Mol Biol. 2021:2291:273-283. doi: 10.1007/978-1-0716-1339-9_12.

Authors

Conor J McGrath¹, Stephanie Schüller²

Affiliations

¹ Norwich Medical School, University of East Anglia, Norwich, UK.
² Norwich Medical School, University of East Anglia, Norwich, UK. s.schuller@uea.ac.uk.

PMID: 33704758
DOI: 10.1007/978-1-0716-1339-9_12

Abstract

The environment in the human intestine is low in oxygen. This affects virulence gene expression of enteropathogens including Shiga toxin-producing E. coli (STEC) and enables mucosal colonization by oxygen-sensitive commensal bacteria. To simulate the oxygen-restricted milieu at the intestinal epithelium, we have developed a vertical diffusion chamber model (VDC) which allows infection of polarized human intestinal epithelia under microaerobic conditions. In this chapter, we present a detailed protocol for performing STEC infections in the VDC system and subsequent analysis of STEC pathogenesis.

Keywords: Host–microbe interactions; Intestinal epithelium; Microaerobiosis; Oxygen; Pathogenesis; Shiga toxin-producing Escherichia coli; Vertical diffusion chamber.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Humans
Intestinal Mucosa* / metabolism
Intestinal Mucosa* / microbiology
Models, Biological*
Oxygen / metabolism*
Shiga-Toxigenic Escherichia coli / metabolism*

Substances

Oxygen

Grants and funding

BB/M011216/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom