Induction of apoptosis in human basophils by anti-Fas antibody treatment in vitro

Int Arch Allergy Immunol. 2008:146 Suppl 1:40-6. doi: 10.1159/000126060. Epub 2008 May 27.

Abstract

Background: Basophils are thought to play an important role in the pathogenesis of allergic inflammation; however, the factors associated with basophil death are not fully understood. Fas (CD95) is a member of the TNF receptor superfamily and is known to induce apoptosis in activated T cells, neutrophils and eosinophils. In the present study, the expression and function of Fas in human basophils were investigated in vitro.

Methods: Human cultured basophils were obtained by culturing cord blood-derived CD34+ cells in the presence of 2.5 ng/ml of IL-3 for 5-6 weeks. The expression of Fas was measured using flow cytometry. Cell viability and morphological changes after the incubation of basophils with anti-Fas mAb (clone CH11, IgM) in the presence of 1 ng/ml of IL-3 were measured using the trypan blue dye exclusion test and light microscopy, respectively.

Results: Human cultured basophils constitutively and significantly expressed Fas on their cell surfaces. Treatment with anti-Fas monoclonal antibody (mAb) significantly reduced basophil viability in a time- and dose-dependent manner. When basophils were incubated with 10 ng/ml of anti-Fas mAb or control for 72 h, the basophil viability was 27.3 +/- 8.8% and 89.3 +/- 5.2%, respectively (p < 0.01). Anti-Fas mAb-treated basophils were shrunken and exhibited condensed nuclei, consistent with apoptosis.

Conclusions: Our findings indicate that human basophils express functional Fas on their cell surfaces, and signaling via Fas may regulate basophil survival in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / pharmacology*
  • Antibodies, Monoclonal, Murine-Derived
  • Apoptosis / immunology*
  • Basophils / drug effects
  • Basophils / immunology*
  • Cell Survival
  • Cells, Cultured
  • Dose-Response Relationship, Immunologic
  • Flow Cytometry
  • Humans
  • fas Receptor / biosynthesis
  • fas Receptor / immunology*

Substances

  • Antibodies, Monoclonal
  • Antibodies, Monoclonal, Murine-Derived
  • anti-Fas monoclonal antibody
  • fas Receptor