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Pathogenesis of immune-mediated cholangiopathy

免疫介导的胆管病的发病机制

基本信息

批准号：
11670473
负责人：
UENO Yoshiyuki
金额：
$ 2.24万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670473/
关键词：
Bile Duct Epithelial Cells Apoptosis Fas (CD95)Fas ligand (FasL)GVHD Fas FasL 胆管炎 TUNEL MRL(lpr lpr)

项目摘要

Bile duct injury observed in hepatic graft versus host disease (GVHD) is regarded as an immune-mediated injury, although its precise mechanism is unclear. However, recent studies have suggested the involvement of Fas-mediated cell death in this immune-mediated cholangiopathy. In this study, we first demonstrated the constitutive expression of Fas receptor by cholangiocytes in situ from normal BALB/c mice, which was upregulated in GVHD mice. Also, we confirmed the Fas protein expression in the isolated cholangiocytes from normal BALB/c mice by immunocytochemistry and immunoblotting. Furthermore, the addition of agonistic Fas antibody (Jo2) induced cholangiocyte apoptosis confirmed by DNA-ladder formation and annexin V staining. Cholangiocytes from Fas-deficient mice (MRL lpr/lpr) did not show Jo2-induced apoptosis. Interferon γ augmented Fas expression and Fas-mediated cell death, respectively. Following these observations, experimental GVHD was induced by transfer of splenocytes from B10.D2 mice to irradiated (800 rad) BALB/c mice. Liver infiltrating lymphocytes from the recipient showed dose-dependent cytotoxicity against ^<51>Cr-labeled cholangiocytes isolated from BALB/c mice. Moreover, the addition of blocking Fas-Fc fusion protein reduced this cytotoxicity to 44.7%. Finally, administration of this Fas-Fc protein to the BALB/c mice, which had been adoptively transferred with splenocytes of B10.D2 mice, prevented the development of hepatic GVHD in vivo. These results demonstrated the involvement of Fas-mediated cell death in cholangiopathy observed in GVHD, and a soluble Fas-Fc protein may have a therapeutic potential for hepatic GVHD.

在肝移植物抗宿主病（GVHD）中观察到的胆管损伤被认为是一种免疫介导的损伤，尽管其确切机制尚不清楚。然而，最近的研究表明，fas介导的细胞死亡参与了这种免疫介导的胆管病。在本研究中，我们首次证实了正常BALB/c小鼠的胆管细胞原位表达Fas受体，该受体在GVHD小鼠中上调。同时，我们通过免疫细胞化学和免疫印迹法证实了正常BALB/c小鼠离体胆管细胞中Fas蛋白的表达。此外，通过dna阶梯形成和膜联蛋白V染色证实，加入激动性Fas抗体（Jo2）可诱导胆管细胞凋亡。fas缺陷小鼠的胆管细胞（MRL lpr/lpr）未显示jo2诱导的凋亡。干扰素γ分别增强Fas表达和Fas介导的细胞死亡。根据这些观察结果，通过转移B10的脾细胞诱导实验性GVHD。D2小鼠与辐照（800 rad） BALB/c小鼠。来自受体的肝浸润淋巴细胞对从BALB/c小鼠分离的^<51> cr标记的胆管细胞表现出剂量依赖性的细胞毒性。此外，阻断Fas-Fc融合蛋白的加入将细胞毒性降低到44.7%。最后，将这种Fas-Fc蛋白给予BALB/c小鼠，BALB/c小鼠已与B10脾细胞过继转移。D2小鼠体内抑制GVHD的发生。这些结果表明，fas介导的细胞死亡参与了GVHD中观察到的胆管病变，可溶性Fas-Fc蛋白可能具有治疗肝GVHD的潜力。