Enterohaemorrhagic, but not enteropathogenic, Escherichia coli infection of epithelial cells disrupts signalling responses to tumour necrosis factor-alpha

Melanie Gareau, Nathan K. Ho, Dirk Brenner, Andrew J. Sousa, Lionel LeBourhis, Tak W. Mak, Stephen E. Girardin, Dana J. Philpott, Philip M. Sherman

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Enterohaemorrhagic Escherichia coli (EHEC), serotype O157: H7 is a non-invasive, pathogenic bacterium that employs a type III secretion system (T3SS) to inject effector proteins into infected cells. In this study, we demonstrate that EHEC blocks tumour necrosis factor-alpha (TNFa)- induced NF-κB signalling in infected epithelial cells. HEK293T and INT407 epithelial cells were challenged with EHEC prior to stimulation with TNFa. Using complementary techniques, stimulation with TNFa caused activation of NF-κB, as determined by luciferase reporter assay (increase in gene expression), Western blotting (phosphorylation of IκBα), immunofluorescence (p65 nuclear translocation) and immunoassay (CXCL-8 secretion), and each was blocked by EHEC O157: H7 infection. In contrast, subversion of host cell signalling was not observed following exposure to either enteropathogenic E. coli, strain E2348/69 (O127: H6) or the laboratory E. coli strain HB101. Heat-killed EHEC had no effect on NF-κB activation by TNFα. Inhibition was mediated, at least in part, by Shiga toxins and by the O157 plasmid, but not by the T3SS or flagellin, as demonstrated by using isogenic mutant strains. These findings indicate the potential for developing novel therapeutic targets to interrupt the infectious process.

Original languageEnglish (US)
Pages (from-to)2963-2973
Number of pages11
JournalMicrobiology
Volume157
Issue number10
DOIs
StatePublished - Oct 2011
Externally publishedYes

Fingerprint

Enterohemorrhagic Escherichia coli
Enteropathogenic Escherichia coli
Escherichia coli Infections
Tumor Necrosis Factor-alpha
Epithelial Cells
Escherichia coli O157
Shiga Toxins
Flagellin
Luciferases
Immunoassay
Fluorescent Antibody Technique
Plasmids
Hot Temperature
Western Blotting
Phosphorylation
Escherichia coli
Bacteria
Gene Expression
Infection
Proteins

ASJC Scopus subject areas

  • Microbiology

Cite this

Enterohaemorrhagic, but not enteropathogenic, Escherichia coli infection of epithelial cells disrupts signalling responses to tumour necrosis factor-alpha. / Gareau, Melanie; Ho, Nathan K.; Brenner, Dirk; Sousa, Andrew J.; LeBourhis, Lionel; Mak, Tak W.; Girardin, Stephen E.; Philpott, Dana J.; Sherman, Philip M.

In: Microbiology, Vol. 157, No. 10, 10.2011, p. 2963-2973.

Research output: Contribution to journalArticle

Gareau, M, Ho, NK, Brenner, D, Sousa, AJ, LeBourhis, L, Mak, TW, Girardin, SE, Philpott, DJ & Sherman, PM 2011, 'Enterohaemorrhagic, but not enteropathogenic, Escherichia coli infection of epithelial cells disrupts signalling responses to tumour necrosis factor-alpha', Microbiology, vol. 157, no. 10, pp. 2963-2973. https://doi.org/10.1099/mic.0.051094-0
Gareau, Melanie ; Ho, Nathan K. ; Brenner, Dirk ; Sousa, Andrew J. ; LeBourhis, Lionel ; Mak, Tak W. ; Girardin, Stephen E. ; Philpott, Dana J. ; Sherman, Philip M. / Enterohaemorrhagic, but not enteropathogenic, Escherichia coli infection of epithelial cells disrupts signalling responses to tumour necrosis factor-alpha. In: Microbiology. 2011 ; Vol. 157, No. 10. pp. 2963-2973.
@article{02011a4aee9c4782a302155c66fcfdf5,
title = "Enterohaemorrhagic, but not enteropathogenic, Escherichia coli infection of epithelial cells disrupts signalling responses to tumour necrosis factor-alpha",
abstract = "Enterohaemorrhagic Escherichia coli (EHEC), serotype O157: H7 is a non-invasive, pathogenic bacterium that employs a type III secretion system (T3SS) to inject effector proteins into infected cells. In this study, we demonstrate that EHEC blocks tumour necrosis factor-alpha (TNFa)- induced NF-κB signalling in infected epithelial cells. HEK293T and INT407 epithelial cells were challenged with EHEC prior to stimulation with TNFa. Using complementary techniques, stimulation with TNFa caused activation of NF-κB, as determined by luciferase reporter assay (increase in gene expression), Western blotting (phosphorylation of IκBα), immunofluorescence (p65 nuclear translocation) and immunoassay (CXCL-8 secretion), and each was blocked by EHEC O157: H7 infection. In contrast, subversion of host cell signalling was not observed following exposure to either enteropathogenic E. coli, strain E2348/69 (O127: H6) or the laboratory E. coli strain HB101. Heat-killed EHEC had no effect on NF-κB activation by TNFα. Inhibition was mediated, at least in part, by Shiga toxins and by the O157 plasmid, but not by the T3SS or flagellin, as demonstrated by using isogenic mutant strains. These findings indicate the potential for developing novel therapeutic targets to interrupt the infectious process.",
author = "Melanie Gareau and Ho, {Nathan K.} and Dirk Brenner and Sousa, {Andrew J.} and Lionel LeBourhis and Mak, {Tak W.} and Girardin, {Stephen E.} and Philpott, {Dana J.} and Sherman, {Philip M.}",
year = "2011",
month = "10",
doi = "10.1099/mic.0.051094-0",
language = "English (US)",
volume = "157",
pages = "2963--2973",
journal = "Microbiology (United Kingdom)",
issn = "1350-0872",
publisher = "Society for General Microbiology",
number = "10",

}

TY - JOUR

T1 - Enterohaemorrhagic, but not enteropathogenic, Escherichia coli infection of epithelial cells disrupts signalling responses to tumour necrosis factor-alpha

AU - Gareau, Melanie

AU - Ho, Nathan K.

AU - Brenner, Dirk

AU - Sousa, Andrew J.

AU - LeBourhis, Lionel

AU - Mak, Tak W.

AU - Girardin, Stephen E.

AU - Philpott, Dana J.

AU - Sherman, Philip M.

PY - 2011/10

Y1 - 2011/10

N2 - Enterohaemorrhagic Escherichia coli (EHEC), serotype O157: H7 is a non-invasive, pathogenic bacterium that employs a type III secretion system (T3SS) to inject effector proteins into infected cells. In this study, we demonstrate that EHEC blocks tumour necrosis factor-alpha (TNFa)- induced NF-κB signalling in infected epithelial cells. HEK293T and INT407 epithelial cells were challenged with EHEC prior to stimulation with TNFa. Using complementary techniques, stimulation with TNFa caused activation of NF-κB, as determined by luciferase reporter assay (increase in gene expression), Western blotting (phosphorylation of IκBα), immunofluorescence (p65 nuclear translocation) and immunoassay (CXCL-8 secretion), and each was blocked by EHEC O157: H7 infection. In contrast, subversion of host cell signalling was not observed following exposure to either enteropathogenic E. coli, strain E2348/69 (O127: H6) or the laboratory E. coli strain HB101. Heat-killed EHEC had no effect on NF-κB activation by TNFα. Inhibition was mediated, at least in part, by Shiga toxins and by the O157 plasmid, but not by the T3SS or flagellin, as demonstrated by using isogenic mutant strains. These findings indicate the potential for developing novel therapeutic targets to interrupt the infectious process.

AB - Enterohaemorrhagic Escherichia coli (EHEC), serotype O157: H7 is a non-invasive, pathogenic bacterium that employs a type III secretion system (T3SS) to inject effector proteins into infected cells. In this study, we demonstrate that EHEC blocks tumour necrosis factor-alpha (TNFa)- induced NF-κB signalling in infected epithelial cells. HEK293T and INT407 epithelial cells were challenged with EHEC prior to stimulation with TNFa. Using complementary techniques, stimulation with TNFa caused activation of NF-κB, as determined by luciferase reporter assay (increase in gene expression), Western blotting (phosphorylation of IκBα), immunofluorescence (p65 nuclear translocation) and immunoassay (CXCL-8 secretion), and each was blocked by EHEC O157: H7 infection. In contrast, subversion of host cell signalling was not observed following exposure to either enteropathogenic E. coli, strain E2348/69 (O127: H6) or the laboratory E. coli strain HB101. Heat-killed EHEC had no effect on NF-κB activation by TNFα. Inhibition was mediated, at least in part, by Shiga toxins and by the O157 plasmid, but not by the T3SS or flagellin, as demonstrated by using isogenic mutant strains. These findings indicate the potential for developing novel therapeutic targets to interrupt the infectious process.

UR - http://www.scopus.com/inward/record.url?scp=80053447743&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80053447743&partnerID=8YFLogxK

U2 - 10.1099/mic.0.051094-0

DO - 10.1099/mic.0.051094-0

M3 - Article

C2 - 21798984

AN - SCOPUS:80053447743

VL - 157

SP - 2963

EP - 2973

JO - Microbiology (United Kingdom)

JF - Microbiology (United Kingdom)

SN - 1350-0872

IS - 10

ER -