title: Ceramide as a TLR4 agonist; a putative signalling intermediate between sphingolipid receptors for microbial ligands and TLR4.
abstract: Mucosal Toll-like receptors (TLRs) respond to pathogens, but remain inert to the indigenous flora, suggesting that the TLRs can receive pathogen-specific signals. For example, TLR4 signalling is activated in CD14-negative epithelial cells by P-fimbriated, uropathogenic Escherichia coli, but not by lipopolysaccharide. The fimbriae use glycosphingolipids as recognition receptors and there is release of ceramide, which is the membrane-anchoring domain of the receptors. In this study, ceramide was identified as a TLR4 agonist and as a putative signalling intermediate between the glycosphingolipid recognition receptors and TLR4. Exogenous ceramide activated a TLR4-dependent epithelial cell response, as shown by exposing stably transfected TLR4-positive or -negative human embryonal kidney cells to C2 and C6 ceramide. A similar, TLR4-dependent response occurred after deliberate release of endogenous long-chained ceramide with sphingomyelinase. Microbial ligands with glycosphingolipid specificity (P fimbriae or the B subunit of Shiga toxin) were shown to increase the levels of ceramide and to trigger a TLR4-dependent response in epithelial cells. The results show that ceramide activates TLR4 signalling and suggest that this mechanism might allow pathogens to elicit mucosal TLR4 responses by perturbing sphingolipid receptors for virulence ligands like P fimbriae.
authors.affiliation: Department of Microbiology, Immunology and Glycobiology, Institute of Laboratory Medicine, Lund University, Sölvegatan 23, S-22362 Lund, Sweden.
authors.forename: Hans
authors.initials: H
authors.lastname: Fischer
authors.forename: Patrik
authors.initials: P
authors.lastname: Ellström
authors.forename: Kristina
authors.initials: K
authors.lastname: Ekström
authors.forename: Lotta
authors.initials: L
authors.lastname: Gustafsson
authors.forename: Mattias
authors.initials: M
authors.lastname: Gustafsson
authors.forename: Catharina
authors.initials: C
authors.lastname: Svanborg
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002518:Ceramides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D016207:Cytokines / Q000378:metabolism; D004187:Disaccharides / Q000737:chemistry / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism / Q000382:microbiology; D004926:Escherichia coli / Q000254:growth & development; D010861:Fimbriae, Bacterial / Q000378:metabolism; D006028:Glycosphingolipids / Q000378:metabolism; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000378:metabolism; D008070:Lipopolysaccharides / Q000378:metabolism / Q000494:pharmacology; D051379:Mice; D018613:Microscopy, Confocal; D015394:Molecular Structure; D015398:Signal Transduction / Q000187:drug effects; D013108:Sphingomyelin Phosphodiesterase / Q000378:metabolism; D013997:Time Factors; D051197:Toll-Like Receptor 4 / Q000819:agonists* / Q000235:genetics / Q000378:metabolism
chemical_list: D002518:Ceramides; D016207:Cytokines; D004187:Disaccharides; D006028:Glycosphingolipids; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D051197:Toll-Like Receptor 4; C032103:4-O-alpha-D-galactopyranosyl-D-galactose; D013108:Sphingomyelin Phosphodiesterase