title: Involvement of lipopolysaccharide binding protein, CD14, and Toll-like receptors in the initiation of innate immune responses by Treponema glycolipids.
abstract: Culture supernatants from Treponema maltophilum associated with periodontitis in humans and Treponema brennaborense found in a bovine cattle disease accompanied with cachexia caused a dose-dependent TNF-alpha synthesis in human monocytes increasing with culture time. This activity could be reduced significantly by blocking the CD14-part of the LPS receptor using the My 4 mAb and by polymyxin B. In the murine macrophage cell line RAW 264.7, Treponema culture supernatants induced TNF-alpha secretion in a LPS binding protein (LBP)-dependent fashion. To enrich for active compounds, supernatants were extracted with butanol, while whole cells were extracted using a phenol/water method resulting in recovery of material exhibiting a similar activity profile. An LPS-LBP binding competition assay revealed an interaction of the treponeme phenol/water extracts with LBP, while precipitation studies implied an affinity to polymyxin B and endotoxin neutralizing protein. Macrophages obtained from C3H/HeJ mice carrying a Toll-like receptor (TLR)-4 mutation were stimulated with treponeme extracts for NO release to assess the role of TLRs in cell activation. Furthermore, NF-kappaB translocation in TLR-2-negative Chinese hamster ovary (CHO) cells was studied. We found that phenol/water-extracts of the two strains use TLRs differently with T. brennaborense-stimulating cells in a TLR-4-dependent fashion, while T. maltophilum-mediated activation apparently involved TLR-2. These results indicate the presence of a novel class of glycolipids in Treponema initiating inflammatory responses involving LBP, CD14, and TLRs.
authors.affiliation: Institut für Mikrobiologie und Hygiene, Universitätsklinikum Charité Medizinische Fakultät der Humboldt-Universität zu Berlin, Berlin, Germany.
authors.forename: N W
authors.initials: NW
authors.lastname: Schröder
authors.forename: B
authors.initials: B
authors.lastname: Opitz
authors.forename: N
authors.initials: N
authors.lastname: Lamping
authors.forename: K S
authors.initials: KS
authors.lastname: Michelsen
authors.forename: U
authors.initials: U
authors.lastname: Zähringer
authors.forename: U B
authors.initials: UB
authors.lastname: Göbel
authors.forename: R R
authors.initials: RR
authors.lastname: Schumann
mesh_terms: D000209:Acute-Phase Proteins; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides; D060829:Arthropod Proteins; D001665:Binding Sites / Q000276:immunology; D001692:Biological Transport / Q000235:genetics; D001769:Blood / Q000382:microbiology; D000440:Butanols; D016466:CHO Cells; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D002460:Cell Line; D002473:Cell Wall / Q000737:chemistry; D002478:Cells, Cultured; D006224:Cricetinae; D016207:Cytokines / Q000096:biosynthesis; D029721:Drosophila Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006017:Glycolipids / Q000276:immunology*; D006801:Humans; D007113:Immunity, Innate; D007447:Invertebrate Hormones / Q000378:metabolism; D018950:Lipopolysaccharide Receptors / Q000502:physiology*; D008070:Lipopolysaccharides / Q000378:metabolism*; D017737:Macrophages, Peritoneal / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D008562:Membrane Glycoproteins / Q000502:physiology*; D051379:Mice; D008809:Mice, Inbred C3H; D009000:Monocytes / Q000276:immunology / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis; D019800:Phenol; D011112:Polymyxin B / Q000378:metabolism / Q000494:pharmacology; D011956:Receptors, Cell Surface / Q000502:physiology*; D011994:Recombinant Proteins / Q000494:pharmacology; D012834:Silver; D013194:Staining and Labeling; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D051193:Toll-Like Receptors; D014208:Treponema / Q000737:chemistry / Q000254:growth & development / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis; D014867:Water; D000099231:Lipopolysaccharide-Binding Protein
chemical_list: D000209:Acute-Phase Proteins; D000911:Antibodies, Monoclonal; D023181:Antimicrobial Cationic Peptides; D060829:Arthropod Proteins; D000440:Butanols; D002352:Carrier Proteins; D016207:Cytokines; D029721:Drosophila Proteins; D006017:Glycolipids; D007447:Invertebrate Hormones; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D008562:Membrane Glycoproteins; D016328:NF-kappa B; D009569:Nitric Oxide; D019800:Phenol; D011112:Polymyxin B; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; D012834:Silver; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha; D014867:Water; D000099231:Lipopolysaccharide-Binding Protein; C495342:TLR2 protein, human; C495345:TLR4 protein, human; C034705:antilipopolysaccharide factor (Limulus)