Critical role for caspases 3 and 8 in neutrophil but not eosinophil apoptosis
Critical role for caspases 3 and 8 in neutrophil but not eosinophil apoptosis
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DOI：
10.1159/000049506
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发表时间：
2001-10-01
影响因子：
2.8
通讯作者：
Simon, HU
中科院分区：
文献类型：
--
作者：
Daigle, I;Simon, HU
背景：细胞凋亡是多细胞生物体控制细胞数量的必要过程。在许多慢性炎性疾病中，不同类型粒细胞的细胞死亡减少是细胞积聚的一个重要机制。在这里，我们研究了半胱天冬酶在中性粒细胞和嗜酸性粒细胞凋亡的存在或不存在的粒细胞-巨噬细胞刺激因子和抗CD 95单克隆抗体，分别的作用。方法：使用标准方案从人血液中分离粒细胞。免疫印迹和功能研究与细胞渗透性特异性肽抑制剂进行分析半胱天冬酶参与。通过RT-PCR、流式细胞术和免疫印迹分析Fas受体和Fas配体的表达。通过溴化乙锭排除试验分析细胞死亡。结果：Caspase 3和Caspase 8在体外参与中性粒细胞凋亡的调控。相反，这两个半胱天冬酶似乎没有发挥重要作用，在嗜酸性粒细胞凋亡的调节。然而，广泛的半胱天冬酶抑制剂VAD阻止嗜酸性粒细胞死亡，表明半胱天冬酶也参与嗜酸性粒细胞的凋亡机制。功能性抑制剂研究表明，caspase 9是至关重要的caspase 3和8的激活，至少在中性粒细胞。相反，中性粒细胞或嗜酸性粒细胞的自发性凋亡不太可能是Fas配体/Fas受体分子相互作用的结果。结论：本研究的数据表明中性粒细胞和嗜酸性粒细胞之间的半胱氨酸蛋白酶的使用存在差异。版权所有（C）2001 S. Karger AG，巴塞尔。
Background: Apoptosis is a necessary process to control cell numbers in multicellular organisms. In many chronic inflammatory diseases, reduced cell death of different types of granulocytes is one important mechanism for cell accumulation. Here, we studied the role of caspases in neutrophil and eosinophil apoptosis in the presence or absence of granulocyte-macrophage stimulating factor and anti-CD95 monoclonal antibodies, respectively. Methods: Granulocytes were isolated from human blood using standard protocols. Immunoblot and functional studies with cell-permeable specific peptide inhibitors were performed to analyze caspase involvement. Fas receptor and Fas ligand expression was analyzed by RT-PCR, flow cytometry, and immunoblotting. Cell death was analyzed by ethidium bromide exclusion test. Results: Caspases 3 and 8 are critically involved in the regulation of neutrophil apoptosis in vitro. In contrast, these two caspases did not appear to play a major role in the regulation of eosinophil apoptosis. However, the broad-range caspase inhibitor VAD prevented eosinophil death, indicating that caspases are also involved within the apoptotic machinery of eosinophils. Functional inhibitor studies suggested that caspase 9 is crucial for both caspase 3 and 8 activation, at least in neutrophils. In contrast, spontaneous apoptosis of neutrophils or eosinophils is unlikely to be the consequence of Fas ligand/Fas receptor molecular interactions. Conclusion: The data of this study indicate differences in the usage of caspases between neutrophils and eosinophils. Copyright (C) 2001 S. Karger AG, Basel.