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.

该项目着重于包膜病毒的分子生物学 包括来自负链病毒（麻疹，， 狂犬病），逆转录病毒（AIDS病毒），致DNA病毒（疱疹病毒）。 这 这项研究的目标是研究病毒的基本分子机制 从病毒入口和组装到病毒基因表达的发病机理， 复制。对这些机制的理解是必要的 长期设计新颖的靶向基因输送系统或基因 具有在基因治疗和疫苗中可能应用的表达系统 发展。我们的计划是发展有缺陷的病毒颗粒 用于神经元或非神经元细胞。到目前为止，我们的研究已经集中 囊泡气孔病毒（VSV），但可以采用基本策略 到其他包膜病毒。 达到这些目标，我们发现病毒组装和细胞病变 VSV很可能是独立事件。 我们还发现高 聚合酶蛋白NS与缺陷干扰的亲和力 粒子基因组确定自动缩水水平。这些都是 了解一个关于病毒发病机理的重要观察结果 范围的结束以及持续的建立和维护 另一端的感染。 对于将病毒靶向特定细胞，我们能够插入A 嵌合HIV受体CD4-VSV包膜糖蛋白分为大多数VSV 颗粒。这些颗粒可以通过 CD4蛋白的抗体。我们也能够插入正常人 CD4分子进入VSV包膜。实验正在进行测试 插入的效率以及病毒组装是否取决于 关于VSV包膜的细胞质尾部区域的存在 蛋白质。我们正在开发一个允许的系统 研究病毒时，研究病毒组装的分子机制 包络蛋白不存在和/或被重组嵌合代替 包膜蛋白。使用离甲酸病毒，我们能够表达VSV n NS和L基因并在不存在的情况下复制有缺陷的病毒基因组 助手病毒。这一发现在我们努力的努力中尤其有希望 产生第一个重组透蛋白病毒。