title: Primary bovine colonic cells: a model to study strain-specific responses to Escherichia coli.
abstract: The parasitic or commensal lifestyle of bacteria in different hosts depends on specific molecular interactions with the respective host species. In vitro models to study intestinal bacteria-host interactions in cattle are not available. Bovine primary colonocyte (PC) cultures were generated from colon crypt explants. Up to day 4 of culture, the vast majority of cells were of epithelial phenotype (i.e., expressed cytokeratin but not vimentin). PCs harboured mRNA specific for Toll-like receptors (TLR) 1, TLR3, TLR4 and TLR6 but not for TLR2, TLR5, TLR7, TLR8, TLR9 and TLR10. Six hours after inoculation of PC cultures with Escherichia coli (E. coli) prototype strains representing different pathovars (enterohaemorrhagic E. coli [EHEC], enteropathogenic E. coli [EPEC], enterotoxic E. coli [ETEC]), bacteria were found attached to the cells. EPEC adhesion was accompanied by intracellular actin accumulation. An attenuated laboratory strain (E. coli K12 C600) and a bovine commensal E. coli strain (P391) both did not adhere. Bacterial or LPS challenge of PC cultures resulted in specific increases in mRNA transcripts for IL-8, GRO-alpha, MCP-1, RANTES, and IL-10. The level of mRNA transcripts for TGF-beta stayed constant, while IL-12 mRNA was not detectable. Short-term cultures of PCs, maintaining epithelial cell properties, interacted with commensal and pathogenic bacteria in a strain-specific manner and have proven to be a useful in vitro model to study the interaction of bacteria with the bovine intestinal mucosa.
authors.affiliation: Institute for Hygiene and Infectious Diseases of Animals, Justus-Liebig-University, D-35392 Giessen, Germany.
authors.forename: Philip Simon
authors.initials: PS
authors.lastname: Bridger
authors.forename: Melanie
authors.initials: M
authors.lastname: Mohr
authors.forename: Ivonne
authors.initials: I
authors.lastname: Stamm
authors.forename: Julia
authors.initials: J
authors.lastname: Fröhlich
authors.forename: Wolfram
authors.initials: W
authors.lastname: Föllmann
authors.forename: Sascha
authors.initials: S
authors.lastname: Birkner
authors.forename: Hannah
authors.initials: H
authors.lastname: Metcalfe
authors.forename: Dirk
authors.initials: D
authors.lastname: Werling
authors.forename: Georg
authors.initials: G
authors.lastname: Baljer
authors.forename: Christian
authors.initials: C
authors.lastname: Menge
mesh_terms: D000818:Animals; D001422:Bacterial Adhesion; D002417:Cattle; D002478:Cells, Cultured; D003106:Colon / Q000166:cytology* / Q000276:immunology / Q000382:microbiology*; D004926:Escherichia coli / Q000472:pathogenicity*; D023421:Models, Animal; D012333:RNA, Messenger / Q000032:analysis; D013045:Species Specificity; D051193:Toll-Like Receptors / Q000235:genetics
chemical_list: D012333:RNA, Messenger; D051193:Toll-Like Receptors