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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

细胞凋亡在支气管哮喘慢性嗜酸性粒细胞炎症的消退中起重要作用。利用人髓母细胞白血病EoL-1细胞系，我们研究了干扰素-γ (IFN-γ)对Fas-和肿瘤坏死因子-α (TNF)诱导的细胞凋亡的影响。THN和抗fas单克隆抗体（CH-11）均可诱导EoL-1细胞凋亡。IFN-γ预处理通过上调Fas增强ch -11诱导的细胞凋亡。然而，治疗明显抑制tnf诱导的细胞凋亡。流式细胞术分析显示，EoL-1表达两种类型的肿瘤坏死因子受体（TNFR1、2），IFN-Γ治疗后，TNFR2表达显著上调，TNFR1表达不上调。tnf诱导的细胞凋亡被TNFR1刺激抗体（htr-9）模拟，并被TNFR1阻断抗体（H398）逆转。虽然tnfr1介导的细胞毒性信号不受IFN-γ预处理的影响，但通过特异性拮抗抗体（utr-1…More）阻断TNFR2可取消IFN-Γ的抑制作用。综上所述，tnf诱导的细胞凋亡是由TNFR1优先介导的，IFN-γ的抗凋亡作用是由上调TNFR2在EoL-1细胞系中的作用引起的。进一步，我们研究了IFN-γ处理的EoL-1细胞中TNFR2的信号转导途径。我们通过western blot分析发现，通过TNFR2的刺激激活了NF-kappaB。最近，我们报道了Rho/ROCK信号通路对细胞功能的影响。Rho/Rock信号抑制肌凝蛋白磷酸酶，使磷酸化的肌凝蛋白保持高水平。ROCK抑制剂（Y-27632）可使麝香型支气管平滑肌收缩放松。肌球蛋白调节细胞的形态变化和迁移。因此，我们研究了ROCK抑制剂（Y-27632）对嗜酸性粒细胞迁移的影响，发现Y-27632抑制嗜酸性粒细胞的迁移，但不抑制Ca γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ γ。这些发现正在准备发表论文。少