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

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. Copyright (C) 2008 S. Karger AG, Basel.