Invasion of bovine epithelial cells by verocytotoxin-producing Escherichia coli O157: H7

Matthews, Karl R.; Murdough, P.A.; Bramley, A. J.

doi:10.1111/j.1365-2672.1997.tb02851.x

Cited by 6 publications

(9 citation statements)

References 14 publications

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“…This binding triggers actin rearrangements and the formation of pedestal structures, representative of A/E lesions. In this study, both O157 : H7 strains were positive by the FAS reaction, confirming that colonization of MAC-T bovine epithelial cells is also mediated through the intimin-Tir interaction, consistent with prior findings (Matthews et al, 1997). However, TW14359 adhered to epithelial cells in significantly greater numbers than Sakai, within 1 h post-infection.…”

Section: Discussionsupporting

confidence: 80%

“…However, TW14359 adhered to epithelial cells in significantly greater numbers than Sakai, within 1 h post-infection. The lack of a difference in invasion levels between TW14359 and Sakai is also in agreement with previous studies (Dibb-Fuller et al, 2001;Matthews et al, 1997). Despite the magnitude of the difference between strains, it is important to note that these data were generated using a monoculture of bovine epithelial cells, which do not accurately represent the complexity of the intestinal environment and the multicellular composition of the mucosal lining in both humans and bovines.…”

Section: Discussionsupporting

confidence: 56%

“…Bovine mammary epithelial (MAC-T) cells (Huynh et al, 1991) are commonly used in studies of adherent and invasive E. coli (Dogan et al, 2006;Dopfer et al, 2000), and E. coli O157 : H7 was previously shown to induce A/E lesions on MAC-T cells (Matthews et al, 1997). MAC-T cells were grown in 75 cm 2 tissue flasks with DMEM [10 % fetal bovine serum (FBS) and 2 % AntibioticAntimycotic solution (Invitrogen)], at 37 uC with 5 % CO 2 .…”

Section: Methodsmentioning

confidence: 99%

“…Propidium iodide counter-staining of specimens confirmed co-localization of actin pedestals and bacteria. Both strains demonstrated the ability to form densely packed microcolonies, or 'localized adherence' (Dibb-Fuller et al, 2001;Matthews et al, 1997;Naylor et al, 2005;Nougayrede et al, 2003), on the surface of MAC-T cells. A/E lesions were not observed following infection with E. coli K-12 (Fig.…”

Section: Interaction Of O157 : H7 With Mac-t Bovine Epithelial Cellsmentioning

confidence: 99%

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Differences in adherence and virulence gene expression between two outbreak strains of enterohaemorrhagic Escherichia coli O157 : H7

et al. 2010

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The Escherichia coli O157 : H7 TW14359 strain was implicated in a multi-state outbreak in North America in 2006, which resulted in high rates of severe disease. Similarly, the O157 : H7 RIMD0509952 (Sakai) strain caused the largest O157 : H7 outbreak to date. Both strains were shown to represent divergent phylogenetic lineages. Here we compared global gene expression patterns before and after epithelial cell exposure, as well as the ability to adhere to and invade epithelial cells, between the two outbreak strains. Epithelial cell assays demonstrated a 2.5-fold greater adherence of the TW14359 strain relative to Sakai, while whole-genome microarrays detected significant differential expression of 914 genes, 206 of which had a fold change ≥1.5. Interestingly, most locus of enterocyte effacement (LEE) genes were upregulated in TW14359, whereas flagellar and chemotaxis genes were primarily upregulated in Sakai, suggesting discordant expression of these genes between the two strains. The Shiga toxin 2 genes were also upregulated in the TW14359 strain, as were several pO157-encoded genes that promote adherence, including type II secretion genes and their effectors stcE and adfO. Quantitative RT-PCR confirmed the expression differences detected in the microarray analysis, and expression levels were lower for a subset of LEE genes before versus after exposure to epithelial cells. In all, this study demonstrated the upregulation of major and ancillary virulence genes in TW14359 and of flagellar and chemotaxis genes in Sakai, under conditions that precede intimate bacterial attachment to epithelial cells. Differences in the level of adherence to epithelial cells were also observed, implying that these two phylogenetically divergent O157 : H7 outbreak strains vary in their ability to colonize, or initiate the disease process.

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Section: Discussionsupporting

confidence: 80%

Section: Discussionsupporting

confidence: 56%

Section: Methodsmentioning

confidence: 99%

Section: Interaction Of O157 : H7 With Mac-t Bovine Epithelial Cellsmentioning

confidence: 99%

See 2 more Smart Citations

Differences in adherence and virulence gene expression between two outbreak strains of enterohaemorrhagic Escherichia coli O157 : H7

et al. 2010

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“…Further studies with a library of O157:H7 isolates of different provenance in adherence and invasion assays are in hand to assess this possibility. Matthews et al [32] demonstrated adherence and invasion of bovine MAC-T mammary secretory and MDBK kidney epithelial cell lines by 12 distinct EHEC O157:H7 strains. Thus, invasion of the bovine cells in this study was anticipated, albeit signi®cantly lower by a factor of 3±4-fold than for S. Typhimurium.…”

Section: Discussionmentioning

confidence: 99%

An in-vitro model for studying the interaction of Escherichia coli O157:H7 and other enteropathogens with bovine primary cell cultures

Dibb-Fuller¹,

Best

Stagg³

et al. 2001

Journal of Medical Microbiology

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Sections of kidney, trachea, ileum, colon, rectum and rumen were removed at post mortem from a neonatal calf and, with the exception of the rumen, primary cell lines were established for each of the cell types. The adherence of enterohaemorrhagic Escherichia coli (EHEC) serotype O157:H7, enteropathogenic E. coli (EPEC) serotype O111, E. coli K12 (a laboratory adapted non-pathogenic strain) and Salmonella enterica serotype Typhimurium was assayed on each cell type. For all adherence assays on all cell lines, EHEC O157:H7 adhered to a signi®cantly greater extent than the other bacteria. S. Typhimurium and EPEC O111 adhered to a similar extent to one another, whereas E. coli K12 was signi®cantly less adherent by 100-fold. In all cell types, >10% of adherent S. Typhimurium bacteria invaded, whereas c. 0.01±0.1% of adherent EHEC O157:H7 and EPEC O111 bacteria invaded, although they are regarded as non-invasive. EHEC O157 generated actin re-arrangements in all cell types as demonstrated by¯uorescent actin staining (FAS) under densely packed bacterial micro-colonies. EPEC O111 readily generated the localised adherent phenotype on bovine cells but generated only densely packed micro-colonies on HEp-2 cells. The intensity of actin re-arrangements induced in bovine cells by EPEC O111 was less than that induced by EHEC O157:H7. The intimate attachment on all cell types by both EHEC O157:H7 and EPEC O111 was clearly demonstrated by scanning electron microscopy.

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Comparison of the sensitivity of manual and automated immunomagnetic separation methods for detection of Shiga toxin-producing Escherichia coli O157:H7 in milk

Reinders¹,

Barna²,

Lipman³

et al. 2002

J Appl Microbiol

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Aim: To determine the sensitivity of methods for detection of injured and uninjured Escherichia coli O157:H7 (E. coli O157) in raw and pasteurized milk. Methods and Results: Raw milk, pasteurized milk with 1AE5% fat content and pasteurized milk with 3AE5% fat content were spiked with E. coli O157 at low levels. The samples were enriched in modified tryptone soya broth with novobiocin (mTSBn) at 37°C. Aliquots of the enriched culture were analysed either by manual immunomagnetic separation (MIMS) and culturing on sorbitol MacConkey agar with or without cefixime and potassium tellurite (SMACct or SMAC), or by automated immunomagnetic separation and integrated ELISA (EiaFoss TM ). Uninjured E. coli O157 organisms were detected in milk by both methods at 1 cfu 10 ml -1 sample). Injured organisms were detected at levels of about 4 cfu 10 ml -1 sample. Direct enrichment in mTSBn (22 h incubation) showed better sensitivity for injured cells than enrichment in buffered peptone water (BPW, 22 h incubation), or in a two-step enrichment consisting of BPW (6 h, 37°C) and mTSBn (16 h, 37°C), successively. Conclusions: The methods showed equal sensitivity in that they were both able to detect 1 cfu 10 ml -1 milk sample. Injured organisms can be detected and isolated at a level almost as low as this. A resuscitation step is not recommended for the detection and isolation of injured and non-injured E. coli O157 from milk. Significance and Impact of the Study: Due to the dilution of contamination in the bulk tank, analysis of milk for the presence of E. coli O157 requires a very sensitive method. Both methods described here are useful for such analysis.

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Invasion of bovine epithelial cells by verocytotoxin-producing Escherichia coli O157: H7

Cited by 6 publications

References 14 publications

Differences in adherence and virulence gene expression between two outbreak strains of enterohaemorrhagic Escherichia coli O157 : H7

Differences in adherence and virulence gene expression between two outbreak strains of enterohaemorrhagic Escherichia coli O157 : H7

An in-vitro model for studying the interaction of Escherichia coli O157:H7 and other enteropathogens with bovine primary cell cultures

Comparison of the sensitivity of manual and automated immunomagnetic separation methods for detection of Shiga toxin-producing Escherichia coli O157:H7 in milk

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