ANALYSIS OF THE PATHOGENESIS OF PRIMARY BILIARY CIRRHOSIS BY USING ANIMAL MODEL THAT IS ESTABLISHED BY MULTI STEP BREAKING OF TOLERANCE

多步突破耐受建立的动物模型分析原发性胆汁性肝硬化的发病机制

• 批准号: 14570512
• 负责人: ZENIYA Mikio
• 金额: $ 2.11万
• 依托单位: JIKEI 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: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570512/
• 关键词: PBC; Animal Model; Dendritic cell; Regulatory cell; 動物モデル; 自己抗原; DC; 調節性細胞; IL-12; 原発性胆汁性肝硬変; PDC-E2

Background : We tried to analyze the pathogenesis of primary biliary cirrhosis(PBC) by using animal model that was established by multi step breakdown of self-tolerance.Methods and Results : <First year> : At first we tried to establish animal model of PBC by inoculating the dendritic cell that was pulsed with recombinant PDC-E2,but bile duct injury was not observed in that model. Next we tried to make the model by inoculating the fusion cell(FC) of dendritic cell and bile duct cell following injection of IL-12. The bile duct injury was observed in that model.<Second year> : We analyzed the immunological condition of established model. The number of intrahepatic Tr cell which produce IL-10 and TGF-B was not changed after FC inoculation. The number of intrahepatic CD25 CD4 positive T reg was slightly decreased after FC inoculation, but the profile of cytokine production of T reg was not changed after FC inoculation. The degree of intrahepatic accumulation of activated T cell was increased after IL-12 inoculation. The degree of the expressions of ICAM-1,VCAM-1 and VAP-1 on sinusoidal endothelial cell was increased after IL-12 inoculation.<Third year> : We tried to identify the auto-antigen in this animal model by using SELEX method, but we could not detect it because of technical problem of experiment.Conclusion : These findings indicate that the decrease of intrahepatic regulatory cell participated in the pathogenesis of PBC.

背景资料：本研究通过自身免疫耐受的多步破坏建立原发性胆汁性肝硬化（PBC）的动物模型，探讨PBC的发病机制。方法和结果<First year>：首先，我们尝试用重组PDC-E2致敏的树突状细胞（DC）构建PBC的动物模型，但未观察到胆管损伤。随后，我们尝试通过接种树突状细胞和胆管细胞的融合细胞（FC）并注射IL-12来制备模型。观察胆管损伤情况。<Second year>对建立的模型进行免疫学分析。接种FC后肝内产生IL-10和TGF-β的Tr细胞数量无明显变化。接种FC后，肝内CD 25、CD 4阳性T reg细胞数略有减少，但T reg细胞因子产生谱无明显变化。IL-12可使肝内活化T细胞聚集程度增加。IL-12刺激后，肝窦内皮细胞ICAM-1、VCAM-1和VAP-1表达增强。<Third year>:本实验采用SELEX方法对该动物模型进行了自身抗原的鉴定，但由于实验技术上的问题，未能检测到自身抗原。结论：肝内调节细胞的减少参与了PBC的发病过程。