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Identification of haptoglobin as an alternative ligand for CD11b/CD18

J Immunol. 1996 Apr 1;156(7):2542-52.

Abstract

Haptoglobin is an acute phase protein with presumed anti-inflammatory activities. We report that purified fluorescein-labeled haptoglobin 1-1 binds to THP1 and U937 promonocytic cell lines, to monocytes, to granulocytes, and to a subset of CD8+ T cells and to NK cells. Studies with radioiodinated haptoglobin on THP1 cells were consistent with specific binding to one class of receptors with a density of 1.7 x 10(5) binding sites per cell and a low affinity of 6.5 x 10(-6) Kd. Binding was increased by Ca2+ and by Ca2+ and ADP. Binding to THP1 and U937 cells could be inhibited by preincubation with nonfluoresceinated haptoglobin and by fibrinogen, but not by albumin, transferrin, or alpha1-acid glycoprotein. Fibrinogen binds to the CD11b/CD18 integrin. We therefore examined whether haptoglobin has the same receptor. The anti-CD11b mAb44 indeed inhibited the binding of fluoresceinated haptoglobin to THP1 and U937 cell lines, and haptoglobin inhibited the binding of the anti-CD11b mAb anti-Leu15 and mAb44 to both cell lines. An anti-CD18 mAb partly inhibited the binding of fluoresceinated haptoglobin to THP1 and U937, indicating that the beta-chain of MAC-1 is also involved in haptoglobin binding. There was no interference between the binding of anti-CD4, anti-CD11a, or anti-CD11c mAb and haptoglobin binding to THP1 cells. Binding of haptoglobin to purified CD11b/CD18 indicates that it binds directly to the receptor. Haptoglobin is an alternative low affinity ligand for the CD11b/CD18 integrin, suggesting that this acute phase protein might regulate MAC-1-dependent cell function in vivo.

MeSH terms

  • Adenosine Diphosphate / pharmacology
  • Antibodies, Monoclonal
  • CD18 Antigens / metabolism*
  • Calcium / pharmacology
  • Cell Line
  • Fibrinogen / metabolism
  • Fluorescein-5-isothiocyanate
  • Fluorescent Dyes
  • Granulocytes / immunology
  • Granulocytes / metabolism
  • Haptoglobins / antagonists & inhibitors
  • Haptoglobins / immunology
  • Haptoglobins / metabolism*
  • Humans
  • In Vitro Techniques
  • Killer Cells, Natural / immunology
  • Killer Cells, Natural / metabolism
  • Kinetics
  • Ligands
  • Lymphocytes / immunology
  • Lymphocytes / metabolism
  • Macrophage-1 Antigen / metabolism*
  • Monocytes / immunology
  • Monocytes / metabolism
  • Protein Binding / drug effects
  • T-Lymphocyte Subsets / immunology
  • T-Lymphocyte Subsets / metabolism
  • Temperature

Substances

  • Antibodies, Monoclonal
  • CD18 Antigens
  • Fluorescent Dyes
  • Haptoglobins
  • Ligands
  • Macrophage-1 Antigen
  • Adenosine Diphosphate
  • Fibrinogen
  • Fluorescein-5-isothiocyanate
  • Calcium