The Study of the mechanisms of fulminant hepatitis B using in vivo hepatitis model.

利用体内肝炎模型研究暴发性乙型肝炎的发病机制。

• 批准号: 06670586
• 负责人: HASEGAWA Kiyoshi
• 金额: $ 0.9万
• 依托单位: Tokyo Women's Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06670586/
• 关键词: hepatitis B virus; fulminant hepatitis; animal model; B型肝炎ウィルス; モデルマウス

In 1993, We analyzed whole sequences of HBV DNA from an patient with fulminant hepatitis B.The result demonstrated that this viral DNA harbared 4,12,15 and 6 missense mutations in envelope, core, pol and X region, respectively.In 1994, in order to study the biological properties of the mutated HBV DNA from fulminant hepatitis B,we transfected the mutated HBV DNA constract into HepG2 cells and checked viral protein in culture medium and viral replication inside the hepatoma cells. As a result, this HBV DNA showed high level of production of HBV core protein and replicative intermediate.In 1995, in order to study how this mutant virus worked in vivo, we established murine hepatitis model, in which viral protein was secreted into circulation and expressed in hepatocytes. We injected the mixture of HBV DNA constract and lipofectin under the capsule of the murine liver. Serial analysis demonstrated that HBsAg was detected in circulation followed by the seroconversion from HBsAg to anti-HBs in circulation and HBsAg and HBcAg was expressed in murine hepatocyte. The study for the establishment of murine fulminant hepatitis model is ongoing.

1993年，我们对1例重型乙型肝炎患者的HBVDNA全序列进行了分析，结果表明，该病毒DNA在包膜、核心区、Pol区和X区分别存在4、12、15和6个错义突变。1994年，我们将突变的HBVDNA转入HepG2细胞，检测培养上清液中的病毒蛋白和肝癌细胞内的病毒复制，以研究其生物学特性。1995年，为了研究这种变异病毒在体内的作用，我们建立了小鼠肝炎模型，病毒蛋白分泌到循环中并在肝细胞中表达。将HBVdna与脂质体的混合物注射到小鼠肝脏包膜下。系列分析表明，在循环中检测到HBs Ag，然后在循环中从HBs Ag到抗-HBs的血清转换，并在小鼠肝细胞中表达了HBs Ag和HBcAg。建立小鼠重型肝炎模型的研究正在进行中。

项目成果

Hasegawa K.et al: "Enhanced replication of a hepatitis B virus mutant associated with an epidemic of fulminant hepatitis." Journal of Virology. 68. 1651-1658 (1994)

Hasekawa K.等人：“与暴发性肝炎流行相关的乙型肝炎病毒突变体的复制增强。”