RRC ID 54145
Author Védrine M, Berthault C, Leroux C, Répérant-Ferter M, Gitton C, Barbey S, Rainard P, Gilbert FB, Germon P.
Title Sensing of Escherichia coli and LPS by mammary epithelial cells is modulated by O-antigen chain and CD14.
Journal PLoS One
Abstract Escherichia coli is one of the major pathogens causing mastitis in dairy cattle. Yet, the factors which mediate the ability for E. coli to develop in the bovine mammary gland remain poorly elucidated. In a mouse model, infections induced by the reference mastitis E. coli P4 showed a strong colonisation of the mammary gland, while this strain had a low stimulating power on cells of the PS bovine mammary epithelial cell line. In order to understand if such a reduced response contributes to the severity of infection, a library of random mutants of P4 strain was screened to identify mutants inducing stronger response of PS cells. Among hyper-stimulating mutants, six were altered in genes involved in biosynthesis of lipopolysaccharide (LPS) and had lost their O-polysaccharide region, suggesting that the presence of O-antigen impairs the response of PS cells to LPS. Using purified smooth (S) and rough (R) fractions of LPS, we showed that the R-LPS fraction induced a stronger response from PS cells than the smooth LPS fraction. Biological activity of the S-LPS fraction could be restored by the addition of recombinant bovine CD14 (rbCD14), indicating a crucial role of CD14 in the recognition of S-LPS by Mammary Epithelial Cells (MEC). When S-LPS and R-LPS were injected in udder quarters of healthy lactating cows, an inflammation developed in all infused quarters, but the S-LPS induced a more intense pro-inflammatory response, possibly in relation to sizeable concentrations of CD14 in milk. Altogether, our results demonstrate that the O-antigen modulates the pro-inflammatory response of MEC to LPS, that S-LPS and R-LPS trigger different responses of MEC and that these responses depend on the presence of CD14.
Volume 13(8)
Pages e0202664
Published 2018-8-24
DOI 10.1371/journal.pone.0202664
PII PONE-D-18-14685
PMID 30142177
PMC PMC6108492
MeSH Animals Cattle Cell Line Chemokines / metabolism Cytokines / metabolism DNA Transposable Elements / genetics Epithelial Cells / cytology Epithelial Cells / metabolism Escherichia coli / growth & development Escherichia coli / isolation & purification Escherichia coli / metabolism* Female HEK293 Cells Humans Lipopolysaccharide Receptors / chemistry Lipopolysaccharide Receptors / genetics Lipopolysaccharide Receptors / metabolism* Lipopolysaccharides / analysis Lipopolysaccharides / immunology* Lipopolysaccharides / metabolism Mastitis, Bovine / immunology Mastitis, Bovine / microbiology Mastitis, Bovine / pathology Milk / metabolism Milk / microbiology Mutagenesis O Antigens / chemistry O Antigens / metabolism* Recombinant Proteins / biosynthesis Recombinant Proteins / chemistry Toll-Like Receptor 2 / metabolism Toll-Like Receptor 4 / metabolism
IF 2.74
Times Cited 3
DNA material pMT-PURO (RDB08532)