ANIMAL MODELS OF HBV: INFECTION AND DISEASE

乙型肝炎动物模型：感染和疾病

• 批准号: 3130279
• 负责人: WILLIAM S ROBINSON
• 金额: $ 36.59万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-01-01 至 1992-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3130279
• 关键词: DNA replication; aflatoxins; animal viral hepatitis; antibody neutralization test; antiviral agents; chemical carcinogen; congenital infection; disease /disorder model; duck hepatitis B virus; electric field; genetic manipulation; hepatitis B virus group; hepatitis virus; hepatocellular carcinoma; hepatotoxin; hybridomas; immunofluorescence technique; molecular cloning; monoclonal antibody; nitrosamines; nucleic acid sequence; oncogenes; squirrel; tissue /cell culture; transfection; transposon /insertion element; virus DNA; virus antigen; virus genetics; virus infection mechanism; virus replication

We propose to continue our studies of disease and infection in 2 animal models of hepatitis B virus, which is associated with hepatitis and hepatocellular carcinoma (HCC) in man. In 1 of the models, ground squirrels infected with ground squirrel hepatitis virus (GSHV), we have seen developed of HCC significantly associated with virus infection, and propose to continue study of the long term effects of virus infection in the animals remaining the colony. We will continue to study the effects of environmental factors such as aflatoxin B1 and antivirals on HCC development. We will determined what common and differentiating characteristics exist between the integrated and unintegrated forms of GSHV DNA in the 8 HCC observed in carrier squirrels and those from studies of WHV- and HBV-related HCC. We will clone and sequence single integrations of GSHV DNA in HCC and study whether c-myc or other oncogenes are rearranged or amplified. We will continue to sequence virus-virus junction areas in the "novel" DNA forms cloned from chronically- infected ground squirrels to determine if these forms reveal a mechanism for integration of viral DNA into chromosomal DNA. In the other animal model, duck hepatitis B virus (DHBV) infection of ducks, we will study what factors influence the type and amount of hepatitis that we have observed in ducklings experimentally injected with DHBV, but absent in congentially- infected animals and uninjected controls. We will follow experimentally-infected ducklings to see if these ducks develop HCC, unlike the congenitally-infected ducklings that we have studied so far. We will use DHBV infection of primary duck hepatocyte cultures to study early events in viral infection, to test monoclonal antibodies to DHBV for neutralizing ability, and to establish a rapid test for antiviral effectiveness. We will attempt to develop alternate cell culture systems for growing DHBV by culturing yolk sac endodermal cells or their precursors and by culturing hepatocytes from ducklings treated with diethylnitrosamine. We will use electroporation-aided transfection of DHBV DNA into both susceptible and non- susceptible cells to study the mechanism of host specificity.

我们建议继续我们的研究疾病和感染在2 B型肝炎病毒的动物模型， 肝炎和肝细胞癌（HCC）。 模型，地松鼠感染了地松鼠肝炎 病毒（GSHV），我们已经看到肝癌的发展显着 与病毒感染有关，并建议继续研究 病毒感染对动物的长期影响 殖民地。 我们会继续研究 环境因素如黄曲霉毒素B1和抗病毒药物对肝癌的影响 发展 我们将决定什么是共同的， 差异化特征存在于综合和 在8例HCC中观察到GSHV DNA的未整合形式， 携带者松鼠和来自WHV和HBV相关研究的松鼠 HCC。 我们将克隆和测序GSHV的单个整合 研究c-myc或其他癌基因是否与肝癌的发生有关。 重新排列或放大。 我们会继续对病毒进行测序 连接区的“新的”DNA形式克隆从慢性- 感染的地松鼠，以确定这些形式是否揭示了 病毒DNA整合到染色体DNA的机制。 在另一种动物模型中，鸭B肝炎病毒（DHBV）感染 的鸭，我们将研究哪些因素影响的类型， 我们在雏鸭中观察到的肝炎数量 实验性注射DHBV，但先天性缺乏- 感染动物和未注射对照。 我们将遵循 实验性感染的小鸭子，看看这些鸭子是否会 肝细胞癌，不像先天性感染的小鸭， 研究到目前为止。 我们将用DHBV感染原代鸭 肝细胞培养研究病毒感染的早期事件， 检测抗DHBV单克隆抗体的中和能力， 建立一种快速检测抗病毒效果的方法。 我们将 尝试开发替代的细胞培养系统， 用卵黄囊内胚层细胞或其前体培养DHBV 并通过培养来自用以下物质处理的雏鸭的肝细胞， 二乙基亚硝胺 我们将使用电穿孔辅助 将DHBV DNA转染易感和非易感细胞， 易感细胞研究宿主特异性的机制。