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目的 探讨Fas单抗对嗜酸细胞 (EOS)凋亡的诱导作用及凋亡EOS的清除机制。方法分离正常人外周血EOS ,加入白细胞介素 (IL) 5与不同浓度 (1～ 10 0 0ng/ml)Fas单抗培养 ,台盼蓝拒染法和末端标记法分别检测细胞存活和凋亡变化 ,观察凋亡EOS能否被巨噬细胞 (MΦ)吞噬。结果 Fas单抗对IL 5介导的EOS存活呈浓度和时间依赖性抑制作用。高浓度的IL 5 (10 6U/L)不能抑制Fas单抗的作用。Fas单抗 (10 0ng/ml)处理 2 4h后EOS即有明显的凋亡 [(35± 6 ) % ],72h后增至 (96±3) % ,前后比较差异有显著性 (P <0 0 1)。Fas单抗 (10 0ng/ml)处理 2 4、48和 72h的EOS与MΦ培养 ,吞噬凋亡EOS的阳性MΦ百分率分别为 (2 0± 5 ) %、(38± 6 ) %和 (4 5± 6 ) % ,MΦ吞噬新分离的EOS阳性百分率为 (1± 2 ) % ,与前者比较差异均有显著性 (P <0 0 1)。结论 Fas抗体能有效诱导EOS凋亡 ,凋亡EOS能被MΦ所吞噬清除
Objective To investigate the induction of apoptosis of eosinophils (EOS) by Fas monoclonal antibody and the mechanism of EOS clearance. Methods Peripheral blood EOS was isolated from healthy volunteers. Interleukin (IL) 5 and Fas monoclonal antibodies (1 ~ 100 ng / ml) were cultured in vitro. Cell viability and apoptosis were detected by trypan blue exclusion and end labeling, respectively Changes, to observe whether apoptotic EOS can be phagocytosed by macrophages (MΦ). Results Fas monoclonal antibody had a concentration-and time-dependent inhibitory effect on IL-5-mediated EOS survival. High concentration of IL 5 (10 6 U / L) did not inhibit the effect of Fas monoclonal antibody. After treated with Fas monoclonal antibody (10 0ng / ml) for 24 hours, EOS showed obvious apoptosis [(35 ± 6)%] and increased to (96 ± 3)% after 72 hours 0 1). The percentages of positive MΦ induced by phagocytosis and apoptosis of EOS and MΦ treated with Fas monoclonal antibody (10 0ng / ml) for 2, 48, and 72h were (20 ± 5)%, (38 ± 6)% and ± 6)%. The positive percentage of EOS phagocytosed newly isolated by MΦ was (1 ± 2)%, which was significantly different from the former (P <0.01). Conclusion Fas antibody can effectively induce apoptosis of EOS, and apoptotic EOS can be phagocytosed by MΦ