West Nile Virus Life Cycle in Flavivirus-Resistant Cells

黄病毒抗性细胞中的西尼罗病毒生命周期

• 批准号: 6954440
• 负责人: PEI-YONG SHI
• 金额: $ 21.15万
• 依托单位: WADSWORTH CENTER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-15 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6954440
• 关键词: West Nile virus; bacterial disease; carbon nitrogen ligase; cell line; genetic susceptibility; laboratory mouse; phenotype; replicon; virus infection mechanism; virus replication

DESCRIPTION (provided by applicant): Host factors play essential roles in modulating viral replication and pathogenesis. The important function of the host factors in flavivirus infections is highlighted by the flavivirus resistance phenotype in mice. Resistant mice can be infected by flaviviruses, but the viral liters in their tissues are 1,000-10,000 times lower than those in tissues of susceptible mice, and the spread of the infections in resistant mice is slower. The resistance to viral-induced morbidity and mortality in mice is flavivirus specific. Genetic studies demonstrated that the flavivirus resistance is controlled by a single locus as an autosomal dominant trait in mice. Recent studies suggested 2'-5'-oligoadenylate synthetase 1b (OAS1b) as a candidate gene for flavivirus-resistance phenotype. Compared with the resistance mice, susceptible mice produce an OAS1b protein lacking 30% of the C-terminal sequence, resulting in the inactivation of the OAS/RNase L pathway. Consequently, a large amount of virus is produced in the susceptible mice. Despite the recent progress, many important questions related to the molecular mechanism of the flavivirus resistance remain unknown. The objective of this application is to explore the molecular details of flavivirus resistance during West Nile virus (WNV) replication. We choose WNV as a model for our study because it is a newly emerging virus in the United States as well as a potential biodefense pathogen. Our recently developed reporting replicons and a full-length infectious clone of WNV has provided us with a unique opportunity to study WNV replication in flavivirus-resistant cells. We will accomplish the overall objective of this application by pursuing two specific aims, (i) Identify the step(s) at which WNV infection is inhibited by the flavivirus resistance gene. (II) Determine viral element(s) genetically associated with the flavivirus resistance gene. A novel approach is taken to identify potential viral element(s) that genetically associates with the flavivirus resistance gene product. Selection of adaptive WNV replicons containing an antibiotic resistance gene in the flavivirus resistant cell line is expected, for the first time, to identify viral elements genetically interacting with the flavivirus resistance gene product. The proposed work is significant, because understanding the molecular mechanism of the flavivirus resistance will provide greater insight into flavivirus replication, pathogenesis, and development of flavivirus vaccine and antiviral therapy.

描述（由申请人提供）：宿主因子在调节病毒复制和发病机制中发挥重要作用。小鼠黄病毒耐药表型凸显了宿主因子在黄病毒感染中的重要作用。耐药小鼠可以被黄病毒感染，但其组织中的病毒量比易感小鼠组织中的病毒量低1000-10000倍，并且耐药小鼠中感染的传播速度较慢。小鼠对病毒引起的发病和死亡的抵抗力是黄病毒特异性的。遗传学研究表明，黄病毒抗性是由单个基因座控制的，作为小鼠的常染色体显性性状。最近的研究表明 2'-5'-寡腺苷酸合成酶 1b (OAS1b) 作为黄病毒抗性表型的候选基因。与耐药小鼠相比，易感小鼠产生的OAS1b蛋白缺少30%的C端序列，导致OAS/RNase L通路失活。因此，易感小鼠体内会产生大量病毒。尽管最近取得了进展，但与黄病毒耐药性分子机制相关的许多重要问题仍然未知。本应用的目的是探索西尼罗河病毒 (WNV) 复制过程中黄病毒耐药性的分子细节。我们选择西尼罗河病毒作为我们研究的模型，因为它是美国新出现的病毒，也是一种潜在的生物防御病原体。我们最近开发的报告复制子和 WNV 的全长感染性克隆为我们提供了研究 WNV 在黄病毒抗性细胞中复制的独特机会。我们将通过追求两个具体目标来实现本申请的总体目标：(i)确定黄病毒抗性基因抑制西尼罗河病毒感染的步骤。 (II) 确定与黄病毒抗性基因遗传相关的病毒元件。采用一种新方法来鉴定与黄病毒抗性基因产物遗传相关的潜在病毒元件。预计首次在黄病毒抗性细胞系中选择含有抗生素抗性基因的适应性WNV复制子，以鉴定与黄病毒抗性基因产物遗传相互作用的病毒元件。这项工作意义重大，因为了解黄病毒耐药的分子机制将为黄病毒复制、发病机制以及黄病毒疫苗和抗病毒治疗的开发提供更深入的了解。