The study of the role of 2 kinds of TNF receptors regulating the TNF-induced apoptosis in eosinophils.

2种TNF受体调节TNF诱导的嗜酸性粒细胞凋亡的作用研究。

基本信息

批准号：
09670466
负责人：
DOBASHI Kunio
金额：
$ 2.37万
依托单位：
Gunma University, School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670466/
关键词：
apoptosis TNF IFN-gannma Eosinophil TNFRI TNFRII EoL-1 Rho ROCK TNF γ-IFN

项目摘要

Apoptosis is important for the resolution of chronic eosinophilic inflammation observed in bronchial asthma. Using a human myeloblastic leukemia cell line, EoL-1, we investigated the effect of interferon-gamma (IFN-γ) on Fas- and tumor becrosis factor-α (TNF)-induced apoptosis. Both THN and anti-Fas monoclonal antibody (CH-11) induced apoptosis of EoL-1 cells. Pretreatment with IFN-γ enhanced the CH-11-induced apoptosis with up-regulation of Fas. However, the treatment markedly inhibited the TNF-induced apoptosis. In flow cytometric analysis, EoL-1 expressed two types of tumor necrosis factor receptors (TNFR1, 2) and the expression of TNFR2 but not of TNFR1 was up-regulated significantly after the IFN-Γ treatment. The TNF-induced apoptosis was mimicked by a TNFR1 stimulating antibody (htr-9), and was reversed by a TNFR1 blocking antibody (H398). Although the TNFR1-mediated cytotoxic signal was not affected by IFN-γ pretreatment, blocking TNFR2 by a specific antagonistic antibody (utr-1 … More ) canceled the inhibitory effect of IFN-Γ. In conclusion, TNF-induced apoptosis was mediated preferentially by TNFR1, and the anti-apoptotic effect of IFN-γ result from up-regulated TNFR2 in EoL-1 cell line. Further, we investigated the signal transduction pathway from TNFR2 in IFN-γ treated EoL-1 cells. We revealed that the stimulation through TNFR2 activated the NF-kappaB by western blot analysis.Recently, we reported that the effect of Rho/ROCK signaling on cell functions. Rho/Rock signaling inhibited the myosin phosphatase and kept the phosphoryrated myosin high. ROCK inhibitor (Y-27632) relaxed the muskarinic bronchiai smooth muscle contraction. Myosin is known to regulates the morphological change and migration of cells. So, we investigated the effect of ROCK inhibitor (Y-27632) on migration of eosinophols and found that Y-27632 inhibitted the migration of eosinophils without inhibitting the CaィイD1++ィエD1 influx or production of active oxygen. These findings were preparing for paper. Less

凋亡对于在支气管哮喘中观察到的慢性嗜酸性感染的分辨率很重要。使用人粒细胞白血病细胞系EOL-1，我们研究了干扰 - 伽马（IFN-γ）对FAS和肿瘤becRosis因子-α（TNF）诱导的细胞凋亡的影响。 THN和抗FAS单克隆抗体（CH-11）均诱导EOL-1细胞凋亡。 IFN-γ的预处理增强了CH-11诱导的凋亡，而Fas的上调。但是，该治疗明显抑制了TNF诱导的细胞凋亡。在流式细胞仪分析中，EOL-1表达了两种类型的肿瘤坏死因子受体（TNFR1、2），而在IFN-γ处理后，TNFR2的表达显着上调了TNFR2。 TNFR1刺激抗体（HTR-9）模仿TNF诱导的凋亡，并通过TNFR1阻断抗体（H398）逆转。尽管TNFR1介导的细胞毒性信号不受IFN-γ预处理的影响，但特定的拮抗抗体（UTR-1…更多）阻止TNFR2取消了IFN-γ的抑制作用。总之，TNFR1优先介导TNF诱导的凋亡，并在EOL-1细胞系中上调TNFR2引起的IFN-γ的抗凋亡作用。此外，我们研究了IFN-γ处理的EOL-1细胞中从TNFR2的信号转移途径。我们透露，通过TNFR2刺激通过Western印迹分析激活了NF-kappab。 RHO/岩石信号传导抑制肌球蛋白磷酸酶，并保持磷酸化的肌球蛋白高。岩石抑制剂（Y-27632）放松了肌肉支气管平滑肌合同。已知肌球蛋白调节细胞的形态变化和迁移。因此，我们研究了岩石抑制剂（Y-27632）对嗜酸酚的迁移的影响，发现Y-27632抑制了嗜酸酚的迁移而不抑制CAIY D1 ++ IE D1 IE D1影响或产生活性氧。这些发现正在准备纸张。较少的