Cell surface binding of LBP-LPS complexes to a protein component distinct from CD14

Richard I. Tapping, Peter S. Tobias

Research output: Contribution to journalArticlepeer-review

Abstract

Lipopolysaccharide (LPS) binding protein (LBP) is a serum molecule that mediates cellular activation in response to endotoxin by ensuring the delivery of LPS to either soluble or membrane bound forms of CD14. Aside from this activating role, previous work has shown that LBP and LPS can bind to cells by forming large aggregates which are anchored by mCD14. This binding phenomenon does not correlate with cellular activation. To further characterize these events, we have generated a biologically active radiolabeled LBP ligand with high specific activity. Through the use of this ligand in whole cell binding assays, we have confirmed that the binding of LBP to CHO cells expressing mCD14 is LPS dependent, blocked by the anti-LBP antibodies 18G4 and 2B5, and appears to involve the self aggregation of LBP-LPS complexes on the cell surface. Moreover, we discovered that non-transfected CHO cells also exhibit a binding phenomenon with all the above characteristics of CHO-mCD14 cells. Binding through this latter receptor(s) is distinct from that mediated by mCD14 in that it is not inhibited by anti-CD14 antibodies 28C5 or 18E12. In addition, unlike binding to mCD14, binding of LBP-LPS complexes to this novel receptor is abolished by pretreatment of cells with trypsin. Using proteinase K we found that LBP-LPS complexes bound either by mCD14 or this new receptor are subsequently internalized. Pretreatment of cells with trypsin also abolishes their ability to internalize mCD14 bound LBP-LPS complexes. The novel receptor for LBP-LPS complexes has been detected on many cell types and may be a receptor required for the cellular clearance of LPS.

Original languageEnglish (US)
Pages (from-to)52-55
Number of pages4
JournalJournal of Endotoxin Research
Volume5
Issue number1-2
DOIs
StatePublished - Jan 1 1999

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Molecular Biology
  • Cell Biology
  • Infectious Diseases

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