Detection of C5a receptors on human eosinophils and inhibition of eosinophil effector functions by anti-C5a receptor (CD88) antibodies

Eur J Immunol. 1996 Jul;26(7):1560-4. doi: 10.1002/eji.1830260723.

Abstract

Eosinophils and complement activation are reported to play a crucial role in the pathogenesis of connective tissue diseases. Depositions of antigens and antigen-antibody complexes lead to complement activation with the generation of anaphylatoxins, particularly C5a, which is thought to be responsible for the infiltration and activation of eosinophils in the tissue. Previous studies suggested that the eosinophil C5a receptor differs structurally from the receptor expressed on neutrophils. In this study, we investigated the expression and functional properties of C5a receptors on human eosinophils using the C5a receptor monoclonal antibody S5/1 (anti-CD88 mAb). Flow cytometric analysis demonstrated that the anti-CD88 mAb bound homogeneously on the surface of human eosinophils from nonatopic healthy donors. In addition, no subpopulations with respect to C5a receptor expression were identified in normodense or hypodense eosinophils of patients with hypereosinophilia. Pre-incubation of eosinophils with anti-CD88 specifically inhibited C5a-induced intracellular calcium concentration transients. C5a-induced chemotactic activity of eosinophils was significantly inhibited after pre-incubation of cells with anti-CD88 mAb in a dose-dependent manner. Furthermore, anti-CD88 mAb inhibited dose-dependently the release of reactive oxygen species by eosinophils following stimulation with C5a. Thus, the human eosinophil C5a receptor is homogeneously expressed on normal eosinophils from healthy donors as well as on hypodense and normodense eosinophil subpopulations from patients with hypereosinophilia. Based on the inhibitory effect of the S5/1 mAb on C5a-stimulated eosinophil effector functions, we conclude that a single C5a receptor type exists on human eosinophils. In addition, the inhibitory effect of the S5/1 mAb on C5a functions may enable a new experimental approach to the treatment of diseases that have been associated with C5a-mediated activation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / pharmacology*
  • Antigens, CD / analysis*
  • Antigens, CD / biosynthesis
  • Antigens, CD / immunology*
  • Calcium / metabolism
  • Chemotaxis, Leukocyte / immunology
  • Complement C5a / antagonists & inhibitors
  • Complement C5a / metabolism*
  • Down-Regulation / drug effects
  • Down-Regulation / immunology
  • Eosinophils / immunology*
  • Eosinophils / metabolism*
  • Humans
  • Intracellular Fluid / immunology
  • Reactive Oxygen Species / metabolism
  • Receptor, Anaphylatoxin C5a
  • Receptors, Complement / analysis*
  • Receptors, Complement / biosynthesis
  • Receptors, Complement / immunology*
  • Tetradecanoylphorbol Acetate / pharmacology

Substances

  • Antibodies, Monoclonal
  • Antigens, CD
  • Reactive Oxygen Species
  • Receptor, Anaphylatoxin C5a
  • Receptors, Complement
  • Complement C5a
  • Tetradecanoylphorbol Acetate
  • Calcium