HEPATITIS VIRUS INFECTION

肝炎病毒感染

• 批准号: 3130277
• 负责人: WILLIAM S ROBINSON
• 金额: $ 38.6万
• 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/3130277
• 关键词: 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; 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.

我们建议在 2 年内继续对疾病和感染进行研究 乙型肝炎病毒动物模型，与 人类肝炎和肝细胞癌（HCC）。 在 1 个 模型，感染地松鼠肝炎的地松鼠 病毒（GSHV），我们已经看到 HCC 显着发展 与病毒感染有关，建议继续研究 病毒感染对剩余动物的长期影响 殖民地。 我们将继续研究其影响 黄曲霉毒素 B1 和抗病毒药物等环境因素对 HCC 的影响 发展。 我们将确定哪些共同点和 集成和集成之间存在差异化特征 在 8 个 HCC 中观察到未整合形式的 GSHV DNA 携带者松鼠以及来自 WHV 和 HBV 相关研究的松鼠 肝癌。 我们将对 GSHV 的单一整合进行克隆和测序 HCC 中的 DNA 并研究 c-myc 或其他癌基因是否 重新排列或放大。 我们将继续对病毒-病毒进行测序 从长期克隆的“新”DNA 形式中的连接区域 受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，鸭乙型肝炎病毒（DHBV）感染 对于鸭子，我们将研究哪些因素影响鸭子的类型和 我们在小鸭身上观察到的肝炎数量 实验性注射 DHBV，但先天性缺失 受感染的动物和未注射的对照。 我们将跟随 实验感染的小鸭子，看看这些鸭子是否发育 HCC，与我们先天感染的小鸭不同 研究到目前为止。 我们将使用DHBV感染的原代鸭 肝细胞培养物研究病毒感染的早期事件 测试 DHBV 单克隆抗体的中和能力，以及 建立抗病毒有效性的快速测试。 我们将 尝试开发替代细胞培养系统以促进生长 通过培养卵黄囊内胚层细胞或其前体细胞进行 DHBV 并通过培养经过处理的小鸭的肝细胞 二乙基亚硝胺。 我们将使用电穿孔辅助 将 DHBV DNA 转染至易感人群和非易感人群中 易感细胞来研究宿主特异性的机制。