REPLICATION AND PATHOGENESIS OF ENVELOPED VIRUSES

有包膜病毒的复制和发病机制

• 批准号: 3881817
• 负责人: M SCHUBERT
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3881817
• 关键词: CD4 molecule; Vesiculovirus; chimeric proteins; gene expression; gene therapy; genetic manipulation; human immunodeficiency virus; interfering virus; phosphoproteins; simian immunodeficiency virus; vaccinia virus; virus envelope; virus infection mechanism; virus protein; virus receptors; virus replication; viruslike particle

This project focuses on the molecular biology of enveloped viruses which include a broad group of viruses from negative strand viruses (measles, rabies), to retroviruses (AIDS virus), to DNA viruses (herpes virus). The goal of this research is to study the basic molecular mechanisms of viral pathogenesis from viral entry and assembly to viral gene expression and replication. The understanding of these mechanisms will be necessary in a long term to design novel targeted gene delivery systems or gene expression systems with possible applications in gene therapy and vaccine development. Our plan is to develop defective virus particles with tropism for neuronal or nonneuronal cells. So far our studies have centered around vesicular stomatitis virus (VSV) but the basic strategies can be applied to other enveloped viruses. Towards these goals we found that viral assembly and cytopathogenesis of VSV are most likely independent events. We also found that the high affinity of the polymerase protein NS to the defective interfering particle genome determines the level of autointerference. These are important observations for the understanding of viral pathogenesis on one end of the spectrum and the establishment and maintenance of persistent infections on the other end. For the targeting of viruses to specific cells we were able to insert a chimeric HIV receptor CD4-VSV envelope glycoprotein into a majority of VSV particles. These particles could be specifically immunoprecipitated with antibodies to the CD4 protein. We were also able to insert normal human CD4 molecule into the VSV envelope. Experiments are in progress to test whether the efficiency of the insertion as well as viral assembly depend on the presence of the cytoplasmic tail region of the VSV envelope protein. We are in the process of developing a system which allows studying the molecular mechanisms of viral assembly when the viral envelope protein is absent and/or replaced by a recombinant chimeric envelope protein. Using vaccinia virus we were able to express the VSV N, NS and L genes and replicate a defective virus genome in the absence of helper virus. This finding is particularly promising in our efforts to generate the first recombinant rhabdovirus.

该项目重点研究包膜病毒的分子生物学 包括负链病毒（麻疹、 狂犬病）、逆转录病毒（艾滋病病毒）、DNA 病毒（疱疹病毒）。 这 本研究的目的是研究病毒的基本分子机制 从病毒进入和组装到病毒基因表达的发病机制 复制。理解这些机制是必要的 长期设计新颖的靶向基因传递系统或基因 表达系统在基因治疗和疫苗中的可能应用 发展。我们的计划是开发具有向性的缺陷病毒颗粒 对于神经元或非神经元细胞。到目前为止我们的研究主要围绕 水疱性口炎病毒（VSV），但可以应用基本策略 对于其他有包膜病毒。 为了实现这些目标，我们发现病毒组装和细胞病变机制 VSV 很可能是独立事件。 我们还发现，高 聚合酶蛋白 NS 对缺陷干扰的亲和力 粒子基因组决定自干扰的水平。这些都是 对于了解病毒发病机制的重要观察 频谱的末端以及持久性的建立和维护 另一端感染。 为了将病毒靶向特定细胞，我们能够插入 嵌合HIV受体CD4-VSV包膜糖蛋白进入大部分VSV 颗粒。这些颗粒可以用以下物质进行特异性免疫沉淀： CD4 蛋白的抗体。我们还能够插入正常人类 CD4 分子进入 VSV 包膜。实验正在进行中以进行测试 插入效率以及病毒组装是否取决于 关于 VSV 包膜细胞质尾区的存在 蛋白质。我们正在开发一个系统，该系统允许 研究病毒组装时的分子机制 包膜蛋白不存在和/或被重组嵌合体取代 包膜蛋白。使用牛痘病毒，我们能够表达 VSV N， NS 和 L 基因并在缺乏的情况下复制有缺陷的病毒基因组 辅助病毒。这一发现对我们的努力特别有希望 产生第一个重组弹状病毒。