An in-vitro model for studying the interaction of Escherichia coli O157:H7 and other enteropathogens with bovine primary cell cultures.
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 significantly greater extent than the other bacteria. S. Typhimurium and EPEC O111 adhered to a similar extent to one another, whereas E. coli K12 was significantly 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 fluorescent 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.
Dibb-Fuller
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MP
Best
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Veterinary Laboratories Agency (Weybridge), Woodham Lane, Addlestone, Surrey KT15 3NB and *Division of Biochemistry and Molecular Biology, School of Biological Sciences, University of Southampton, Bassett Crescent East, Southampton SO16 7PX.
Stagg
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DA
Cooley
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Woodward
M J
MJ
D000818:Animals; D001422:Bacterial Adhesion; D002417:Cattle; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D004064:Digestive System / Q000166:cytology / Q000382:microbiology* / Q000648:ultrastructure; D004926:Escherichia coli / Q000502:physiology / Q000648:ultrastructure; D019453:Escherichia coli O157 / Q000502:physiology* / Q000648:ultrastructure; D007668:Kidney / Q000166:cytology / Q000382:microbiology* / Q000648:ultrastructure; D008854:Microscopy, Electron / Q000662:veterinary; D008855:Microscopy, Electron, Scanning / Q000662:veterinary; D008856:Microscopy, Fluorescence / Q000662:veterinary; D012486:Salmonella typhimurium / Q000502:physiology / Q000648:ultrastructure; D014132:Trachea / Q000166:cytology / Q000382:microbiology* / Q000648:ultrastructure