title: P fimbriae-dependent, lipopolysaccharide-independent activation of epithelial cytokine responses.
abstract: Cells in the mucosal barrier are equipped to sense and respond to microbes in the lumen and translate this molecular information into signals that can reach local or distant sites. The interaction of P-fimbriated Escherichia coli with human uroepithelial cells is a model to study the molecular mechanism of epithelial cell activation by mucosal pathogens. Here, we examine the role of lipopolysaccharide (LPS) as a co-stimulatory molecule in epithelial cell activation by P-fimbriated E. coli. P-fimbriated clinical isolates or recombinant strains were shown to trigger a fimbriae-dependent epithelial cell cytokine response. Mutational inactivation of the msbB sequences that control lipid A myristoylation drastically impaired monocyte stimulation but not epithelial responses to P-fimbriated bacteria. Polymyxin B or bactericidal/permeability increasing factor (BPI) neutralized the effects of lipid A in the monocyte assay, but did not reduce epithelial responses. Finally, isolated LPS of the smooth, rough and deep rough chemotypes were poor epithelial cell activators. The cells were shown to lack surface CD14 or CD14 mRNA as well as the CD14 co-receptor function and were also very poor LPS responders in the presence of human serum. These results demonstrate that epithelial cell responses to P-fimbriated E. coli are CD14 and LPS independent, and suggest that attaching pathogens can overcome the LPS unresponsiveness of epithelial cells by fimbriae-dependent activation mechanisms.
authors.affiliation: Department of Laboratory Medicine, Division of Microbiology, Immunology and Glycobiology, University of Lund, Sölvegatan 23, S-223 62 Lund, Sweden. maria.hedlund@mig.lu.se
authors.forename: M
authors.initials: M
authors.lastname: Hedlund
authors.forename: C
authors.initials: C
authors.lastname: Wachtler
authors.forename: E
authors.initials: E
authors.lastname: Johansson
authors.forename: L
authors.initials: L
authors.lastname: Hang
authors.forename: J E
authors.initials: JE
authors.lastname: Somerville
authors.forename: R P
authors.initials: RP
authors.lastname: Darveau
authors.forename: C
authors.initials: C
authors.lastname: Svanborg
mesh_terms: D000217:Acyltransferases; D023181:Antimicrobial Cationic Peptides; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001798:Blood Proteins / Q000494:pharmacology; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins; D010861:Fimbriae, Bacterial / Q000235:genetics / Q000378:metabolism*; D005838:Genotype; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D008050:Lipid A / Q000031:analogs & derivatives / Q000235:genetics; D018950:Lipopolysaccharide Receptors / Q000235:genetics / Q000276:immunology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008565:Membrane Proteins; D009154:Mutation; D010641:Phenotype; D011112:Polymyxin B / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D012475:Salmonella; D013997:Time Factors
chemical_list: D023181:Antimicrobial Cationic Peptides; D001426:Bacterial Proteins; D001798:Blood Proteins; D016207:Cytokines; D029968:Escherichia coli Proteins; D015850:Interleukin-6; D016209:Interleukin-8; D008050:Lipid A; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D008565:Membrane Proteins; D012333:RNA, Messenger; C021623:bactericidal permeability increasing protein; D000217:Acyltransferases; C097895:MsbB protein, E coli; D011112:Polymyxin B