Lipopolysaccharide binding protein binds to triacylated and diacylated lipopeptides and mediates innate immune responses

J Immunol. 2004 Aug 15;173(4):2683-91. doi: 10.4049/jimmunol.173.4.2683.

Abstract

LPS binding protein (LBP) is an acute-phase protein synthesized predominantly in the liver of the mammalian host. It was first described to bind LPS of Gram-negative bacteria and transfer it via a CD14-enhanced mechanism to a receptor complex including TLR-4 and MD-2, initiating a signal transduction cascade leading to the release of proinflammatory cytokines. In recent studies, we found that LBP also mediates cytokine induction caused by compounds derived from Gram-positive bacteria, including lipoteichoic acid and peptidoglycan fragments. Lipoproteins and lipopeptides have repeatedly been shown to act as potent cytokine inducers, interacting with TLR-2, in synergy with TLR-1 or -6. In this study, we show that these compounds also interact with LBP and CD14. We used triacylated lipopeptides, corresponding to lipoproteins of Borrelia burgdorferi, mycobacteria, and Escherichia coli, as well as diacylated lipopeptides, corresponding to, e.g., 2-kDa macrophage activating lipopeptide of Mycoplasma spp. Activation of Chinese hamster ovary cells transfected with TLR-2 by both lipopeptides was enhanced by cotransfection of CD14. Responsiveness of human mononuclear cells to these compounds was greatly enhanced in the presence of human LBP. Binding of lipopeptides to LBP as well as competitive inhibition of this interaction by LPS was demonstrated in a microplate assay. Furthermore, we were able to show that LBP transfers lipopeptides to CD14 on human monocytes using FACS analysis. These results support that LBP is a pattern recognition receptor transferring a variety of bacterial ligands including the two major types of lipopeptides to CD14 present in different receptor complexes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acute-Phase Proteins / immunology*
  • Acute-Phase Proteins / metabolism*
  • Acylation
  • Animals
  • Antigens, Bacterial / immunology
  • Antigens, Viral / immunology
  • CHO Cells
  • Carrier Proteins / immunology
  • Carrier Proteins / metabolism*
  • Cells, Cultured
  • Cricetinae
  • Flow Cytometry
  • Humans
  • Immunity, Innate
  • Leukocytes, Mononuclear / immunology
  • Lipopolysaccharide Receptors / genetics
  • Lipopolysaccharide Receptors / immunology*
  • Lipopolysaccharide Receptors / metabolism
  • Lipoproteins / chemistry
  • Lipoproteins / immunology*
  • Membrane Glycoproteins / genetics
  • Membrane Glycoproteins / immunology
  • Membrane Glycoproteins / metabolism*
  • Peptides / chemistry
  • Peptides / immunology*
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / immunology
  • Receptors, Cell Surface / metabolism
  • Toll-Like Receptor 1
  • Toll-Like Receptor 2
  • Toll-Like Receptor 4
  • Toll-Like Receptors
  • Transfection

Substances

  • Acute-Phase Proteins
  • Antigens, Bacterial
  • Antigens, Viral
  • Carrier Proteins
  • Lipopolysaccharide Receptors
  • Lipoproteins
  • Membrane Glycoproteins
  • Peptides
  • Receptors, Cell Surface
  • TLR2 protein, human
  • TLR4 protein, human
  • Toll-Like Receptor 1
  • Toll-Like Receptor 2
  • Toll-Like Receptor 4
  • Toll-Like Receptors
  • lipopolysaccharide-binding protein