The role of CD14 in signaling mediated by outer membrane lipoproteins of Borrelia burgdorferi.
Borrelia burgdorferi possesses membrane lipoproteins that exhibit stimulatory properties and, consequently, have been implicated in the pathology related to Lyme disease. As CD14 has been shown to mediate signaling by a number of lipid-modified bacterial products, the involvement of CD14 in signaling mediated by two B. burgdorferi lipoproteins, outer surface protein A (OspA) and OspC, was determined. Lipoprotein-mediated induction of nuclear factor-kappaB nuclear translocation and production of IL-8 and IL-6 in HUVEC was enhanced in the presence of serum or soluble rCD14. CD14-specific Abs that block LPS-mediated signaling also inhibited lipoprotein-dependent signaling in HUVEC and neutrophils. The formation of stable complexes between OspA and CD14 was demonstrated by native gel electrophoresis. LPS was found to compete with OspA for binding with CD14, suggesting that LPS and OspA bind similar regions on CD14. The similarity in binding was further supported by the finding that a mutant soluble CD14, lacking the LPS binding site, did not facilitate lipoprotein signaling, nor did it form a complex with OspA. Binding of OspA to CD14 was dependent on the lipid modification, as unlipidated OspA did not form a complex with CD14 or stimulate cells. In contrast, the lipopeptide remaining after proteinase K digestion both formed a complex with CD14 and retained stimulatory properties. These findings indicate that CD14 facilitates bacterial lipoprotein signaling in mammalian cells.
Wooten
R M
RM
Department of Pathology, University of Utah School of Medicine, Salt Lake City 84132, USA.
Morrison
T B
TB
Weis
J H
JH
Wright
S D
SD
Thieringer
R
R
Weis
J J
JJ
D000276:Adjuvants, Immunologic / Q000502:physiology; D000954:Antigens, Surface / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D001428:Bacterial Vaccines; D001667:Binding, Competitive / Q000276:immunology; D015748:Borrelia burgdorferi Group / Q000276:immunology*; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004355:Drug Stability; D004730:Endothelium, Vascular / Q000166:cytology; D006801:Humans; D007106:Immune Sera / Q000494:pharmacology; D018950:Lipopolysaccharide Receptors / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008074:Lipoproteins / Q000037:antagonists & inhibitors / Q000502:physiology; D046911:Macromolecular Substances; D011485:Protein Binding / Q000276:immunology; D015398:Signal Transduction / Q000276:immunology*; D014471:Umbilical Veins
D000276:Adjuvants, Immunologic; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D016895:Culture Media, Serum-Free; D007106:Immune Sera; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D008074:Lipoproteins; D046911:Macromolecular Substances; C072045:OspA protein