Primary bovine colonic cells: a model to study strain-specific responses to Escherichia coli.
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.
Bridger
Philip Simon
PS
Institute for Hygiene and Infectious Diseases of Animals, Justus-Liebig-University, D-35392 Giessen, Germany.
Mohr
Melanie
M
Stamm
Ivonne
I
Fröhlich
Julia
J
Föllmann
Wolfram
W
Birkner
Sascha
S
Metcalfe
Hannah
H
Werling
Dirk
D
Baljer
Georg
G
Menge
Christian
C
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
D012333:RNA, Messenger; D051193:Toll-Like Receptors