TY - JOUR
T1 - Induction of apoptosis in human basophils by anti-Fas antibody treatment in vitro.
AU - Matsumoto, Kenji
AU - Maeda, Akihiko
AU - Bochner, Bruce S.
AU - Wakiguchi, Hiroshi
AU - Saito, Hirohisa
PY - 2008
Y1 - 2008
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=48849099993&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=48849099993&partnerID=8YFLogxK
U2 - 10.1159/000126060
DO - 10.1159/000126060
M3 - Article
C2 - 18504406
AN - SCOPUS:48849099993
SN - 1018-2438
VL - 146 Suppl 1
SP - 40
EP - 46
JO - International archives of allergy and immunology
JF - International archives of allergy and immunology
ER -