Mechanism of Siglec-8-mediated cell death in IL-5-activated eosinophils: role for reactive oxygen species-enhanced MEK/ERK activation.
Mechanism of Siglec-8-mediated cell death in IL-5-activated eosinophils: role for reactive oxygen species-enhanced MEK/ERK activation.
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DOI：
10.1016/j.jaci.2013.03.024
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发表时间：
2013-08
影响因子：
14.2
通讯作者：
Zimmermann, Nives
中科院分区：
文献类型：
--
作者：
Kano, Gen;Almanan, Maha;Bochner, Bruce S.;Zimmermann, Nives
siglece -8在人嗜酸性粒细胞上表达，其结扎可诱导细胞死亡。矛盾的是，siglece -8介导的细胞死亡在激活和存活因子IL-5的存在下显着增强，并且变得独立于caspase活性。在本报告中，我们研究了活化的嗜酸性粒细胞中siglece -8介导的细胞死亡机制。用激动性抗siglece -8抗体和IL-5处理人外周血嗜酸性粒细胞，流式细胞术和形态学检测细胞死亡情况。phospho-luminex、流式细胞术和Western blotting检测MAPK的磷酸化水平。采用二氢膦胺（DHR）荧光法测定ROS积累。与单独使用抗siglece -8刺激相比，抗siglece -8和IL-5联合刺激显著增加了细胞坏死伴随颗粒释放死亡的速率和比例，其中以凋亡为主。这些发现表明，在抗siglece -8/IL-5共刺激过程中，激活了一种特定的、独特的细胞死亡生化途径。与单独IL-5刺激的细胞相比，co42刺激的细胞中ERK1/2和MEK1的磷酸化显著增强并持续；单独抗siglece -8不引起ERK1/2磷酸化。MEK1抑制剂阻断抗siglece -8/ il -5诱导的细胞死亡。siglece -8连接以不依赖mek的方式诱导ROS积累。相反，ROS抑制剂可阻止抗siglece -8/ il -5诱导的ERK磷酸化增强和细胞死亡。外源性ROS模拟了抗siglece -8的刺激，足以诱导il -5处理细胞的细胞死亡。总的来说，这些数据表明ERK磷酸化的增强是ROS生成的下游。在活化的嗜酸性粒细胞中，siglece -8的连接导致ros依赖性il -5诱导的ERK磷酸化增强，从而导致一种新的生化调节的嗜酸性粒细胞死亡模式。
Siglec-8 is expressed on human eosinophils, where its ligation induces cell death. Paradoxically, Siglec-8-mediated cell death is markedly enhanced by the presence of the activation and survival factor IL-5 and becomes independent of caspase activity. In this report we investigate the mechanism of Siglec-8-mediated cell death in activated eosinophils. Human peripheral blood eosinophils were treated with agonistic anti-Siglec-8 antibody and IL-5, and cell death was determined by flow cytometry and morphology. Phosphorylation of MAPK was determined by phospho-luminex, flow cytometry, and Western blotting. ROS accumulation was determined by dihydrorhodamine (DHR) fluorescence. Co-stimulation with anti-Siglec-8 and IL-5 significantly increased the rate and proportion of cells dying by necrosis accompanied by granule release as compared to stimulation with anti-Siglec-8 alone, in which apoptosis predominated. Together with the caspase-independent mode of cell death in co-stimulated cells, these findings suggest the activation of a specific and distinct biochemical pathway of cell death during anti-Siglec-8/IL-5 co-stimulation. Phosphorylation of ERK1/2 and MEK1 was significantly enhanced and sustained in co42 stimulated cells compared to cells stimulated with IL-5 alone; anti-Siglec-8 alone did not cause ERK1/2 phosphorylation. MEK1 inhibitors blocked anti-Siglec-8/IL-5-induced cell death. ROS accumulation was induced by Siglec-8 ligation in a MEK-independent manner. In contrast, ROS inhibitor prevented the anti-Siglec-8/IL-5-induced enhancement of ERK phosphorylation and cell death. Exogenous ROS mimicked stimulation by anti-Siglec-8 and was sufficient to induce enhanced cell death in IL-5-treated cells. Collectively, these data suggest that the enhancement of ERK phosphorylation is downstream of ROS generation. In activated eosinophils, ligation of Siglec-8 leads to ROS-dependent enhancement of IL-5-induced ERK phosphorylation, which results in a novel mode of biochemically-regulated eosinophil cell death.
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影响因子：
5.5
作者：
Lundqvist-Gustafsson, H;Bengtsson, T
通讯作者：
Bengtsson, T
影响因子：
7.7
作者：
Devin, A;Lin, Y;Liu, ZG
通讯作者：
Liu, ZG
DOI：
10.1111/j.1365-2222.2008.03173.x
发表时间：
2009-03
期刊：
Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology
影响因子：
--
作者：
Bochner BS
通讯作者：
Bochner BS
影响因子：
14.2
作者：
Andina, Nicola;Didichenko, Svetlana;Simon, Hans-Uwe
通讯作者：
Simon, Hans-Uwe
影响因子：
3.7
作者：
Lee, YJ;Cho, HN;Lee, YS
通讯作者：
Lee, YS