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Reovirus Type 3 as a Causative Agent of Biliary Atresia- Immunocytochemical and Molecular Biological Study

3 型呼肠孤病毒作为胆道闭锁的病原体 - 免疫细胞化学和分子生物学研究

基本信息

批准号：
01570546
负责人：
MATSUI Akira M. D.
金额：
$ 0.83万
依托单位：
Jichi Medical School, Faculty of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

项目摘要

The first aim of the present study was to identify reovirus type 3 (Reo 3) antigens and RNAs from the liver tissues of patients with biliary atresia. Immunofluorescence and immunoperoxidase suggested that Reo 3 antigens were localized at cytoplasm of hepatocytes and/or ductular epithelia around the portal tracts in all of 25 patients with biliary atresia. They were negative in 6 of 8 controls with liver disease and all of 3 controls without liver disease. Immunoelectron microscopy suggested that Reo 3 antigens were located at small vesicles in lysosome-like vacuoles of these liver cells. The oligomer M1 probes were designed, synthesized and used for polymerase chain reaction for Reo 3 M1 in those livers. M1 segments were detected in all of 3 livers of biliary atresia, but not in 2 controls suggesting that at least a part of Reo 3 were replicated in these livers. It was thus concluded that our patients with biliary atresia were infected with Reo 3.The second aim of our study was to produce experimental models of biliary atresia in suckling animals and compare the pathogenesis with that of humans. We found Reo 3 antigens in the livers of weanling mice which were infected with Reo 3, but not in the porta hepatis so far. This study should be carried on from now.

本研究的第一个目的是从胆道闭锁患者的肝组织中鉴定呼肠孤病毒3型（Reo 3）抗原和RNA。免疫荧光和免疫过氧化物酶染色显示，25例胆道闭锁患者的Reo 3抗原均定位于汇管区周围的肝细胞和（或）小管上皮细胞的胞浆内。8例肝病对照中6例阴性，3例非肝病对照全部阴性。免疫电镜显示Reo 3抗原定位于肝细胞溶酶体样空泡的小泡内。设计、合成寡聚体M1探针并用于这些肝脏中Reo 3 M1的聚合酶链反应。3例胆道闭锁患者的肝脏中均检测到M1段，而2例对照组中未检测到M1段，表明Reo 3至少有一部分在这些肝脏中复制。本研究的第二个目的是建立哺乳动物胆道闭锁的实验模型，并与人类胆道闭锁的发病机制进行比较。在Reo 3感染的断奶小鼠肝脏中发现了Reo 3抗原，但在肝门区尚未发现。这项研究应该从现在开始进行。