ANIMAL MODELS OF HBV: INFECTION AND DISEASE

我们建议在2年内继续我们对疾病和感染的研究 乙肝病毒的动物模型，这与 人类的肝炎和肝细胞癌。在其中的1个 模型，感染了沙鼠肝炎的沙鼠 病毒(GSHV)，我们已经看到肝癌的发展显著 与病毒感染有关，并建议继续研究 病毒感染对剩余动物的长期影响 殖民地。我们将继续研究这些措施的影响 黄曲霉毒素B1和抗病毒药物等环境因素对肝细胞癌的影响 发展。我们将确定什么是共同的和 整合型和整合型之间存在区别特征 年观察到的8例肝细胞癌中GSHV DNA的非整合形式 携带者松鼠和来自WHV和乙肝相关研究的松鼠 肝细胞癌。我们将对GSHV的单一整合进行克隆和测序 并研究c-myc或其他癌基因是否 重新排列或放大的。我们将继续对病毒-病毒进行排序 从长期克隆的新的DNA形式中的连接区- 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，鸭乙肝病毒(DHBV)感染 在鸭子方面，我们会研究哪些因素会影响鸭子的类型和 我们在雏鸭身上观察到的肝炎的数量 实验中注射了D乙肝病毒，但在同种情况下缺失- 被感染的动物和未注射的对照组。我们会跟随 实验感染的小鸭子，看看这些鸭子是否会发育 肝细胞癌，不像我们有的先天感染的鸭子 目前为止已经研究过了。我们将使用原代鸭的DHBVi感染 肝细胞培养以研究病毒感染的早期事件，以 检测抗DHBV单抗的中和能力，以及 建立一种快速检测抗病毒效果的方法。我们会 尝试开发替代细胞培养系统以供生长 卵黄囊内皮细胞或其前体细胞培养产生的乙肝病毒 并通过培养处理过的雏鸭的肝细胞 二乙基亚硝胺。我们将使用电穿孔辅助 DHBVDNA在易感和非易感人群中的表达 易感细胞研究宿主特异性的机制。

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.