Virus and Host Factors in HSV Cell-Cell Spread

HSV 细胞间传播中的病毒和宿主因素

• 批准号: 8532474
• 负责人: RICHARD J ROLLER
• 金额: $ 37.34万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8532474
• 关键词: Antibody Formation; Area; Capsid; Cell Membrane Proteins; Cell Separation; Cell surface; Cells; Cellular Membrane; Chickenpox; Disease; Dominant-Negative Mutation; E-Cadherin; Epithelial Cells; Exposure to; Genetic Suppression; Glycoproteins; Goals; Golgi Apparatus; Heart; Herpes Simplex Infections; Herpes zoster disease; Herpesviridae; Human; Immune response; Infection; Integration Host Factors; Intercellular Junctions; Life; Mediator of activation protein; Membrane Proteins; Neurons; Nuclear; Pathway interactions; Persons; Play; Process; Property; Proteins; Proteomics; Recurrence; Role; Sensory Ganglia; Simplexvirus; Site; Sorting - Cell Movement; Stratified Epithelium; Surface; Symptoms; Testing; Viral; Viral Genes; Viral Proteins; Virion; Virus; Virus Assembly; cell type; extracellular; ganglion cell; gene function; inhibitor/antagonist; mutant; therapeutic target; tool; trafficking

DESCRIPTION (provided by applicant): All of the manifestations of alphaherpesvirus disease (herpes simplex, varicella zoster) result from the ability of the virus to spread from the initial infected cell or cells at mucosal surfaces to other cells in the area and to nerve cells that innervate the site of primary replication. Recurrence of symptoms and consequent spread of the virus to new hosts similarly requires the ability to spread from the sensory ganglion cells to cell at the periphery and among the cells on the mucosal surface. Amazingly, spread of the virus in recurrent infection occurs in the face of an adaptive immune response, including an antibody response that should neutralize virus released from the cell. The disease-causing properties of these viruses therefore depend on the mechanisms used for spread from cell to cell that protect the virus from exposure to effectors of the adaptive immune response. Spread of the human alphaherpesviruses within the host requires trafficking of newly assembled virus particles from their assembly site at the Golgi to exposed cell surfaces for release to extracellular medium or to cell junctions for cell-to-cell spread (CCS). Neither trafficking pathway is well understood. In part this is because, other than viral proteins required for entry of virus into host cells, no virl gene functions have been identified that are required for the process in most cell types. We have discovered that two viral gene products, pUL34 and pUL51, play critical roles in efficient virus release and/or CCS. Both proteins are apparently multifunctional. pUL34 is required for nuclear egress of herpesvirus capsids, and pUL51 has been shown to be required for efficient cytoplasmic assembly of the virus. We have discovered, however, that both proteins play critical roles in release and CCS that can be genetically uncoupled from their roles in virion assembly. Our overall goal is to test the hypothesis that HSV release and CCS are accomplished by viral hijacking of cellular pathways that sort host cell membrane proteins to appropriate surfaces and junctions. We will use two general approaches to this overall goal. The first approach (contained in the first two specific aims) is to characterize the functions and interactions of pUL34 and pUL51 that are required for virus release and CCS. These viral proteins can thus be used as tools to identify critical viral and cellular proteins that also participate. The second approach (contained in the third specific aim is to take advantage of recently developed information about the host pathways that deliver cellular membrane proteins to basolateral and junctional surfaces of cells and to probe those pathways using dominant negative inhibitors of critical molecules.

描述（由申请方提供）：α疱疹病毒病（单纯疱疹、水痘带状疱疹）的所有表现都是由于病毒能够从粘膜表面的初始感染细胞扩散到该区域的其他细胞以及支配初次复制部位的神经细胞。症状的复发和随后病毒向新宿主的传播同样需要从感觉神经节细胞向周围细胞和粘膜表面细胞之间传播的能力。令人惊讶的是，病毒在反复感染中的传播是在适应性免疫反应的情况下发生的，包括应该中和从细胞释放的病毒的抗体反应。因此，这些病毒的致病特性取决于用于从细胞到细胞传播的机制，该机制保护病毒免于暴露于适应性免疫应答的效应物。 人α疱疹病毒在宿主内的传播需要将新组装的病毒颗粒从其在高尔基体的组装位点运输到暴露的细胞表面以释放到细胞外介质或细胞连接处以进行细胞间传播（CCS）。这两种贩运途径都没有得到很好的了解。在 这部分是因为除了病毒进入宿主细胞所需的病毒蛋白质之外，还没有鉴定出大多数细胞类型中该过程所需的Vir 1基因功能。我们已经发现两种病毒基因产物pUL 34和pUL 51在有效的病毒释放和/或CCS中起关键作用。这两种蛋白质显然是多功能的。pUL 34是疱疹病毒衣壳的核出口所需的，并且已经显示pUL 51是病毒的有效胞质组装所需的。然而，我们已经发现，这两种蛋白质在释放和CCS中起着关键作用，可以从它们在病毒体组装中的作用中遗传解耦。 我们的总体目标是测试这样的假设：单纯疱疹病毒的释放和捕获捕获是通过病毒劫持细胞途径来实现的，这些细胞途径将宿主细胞膜蛋白分类到适当的表面和连接处。我们将使用两种方法来实现这一总体目标。第一种方法（包含在前两个具体目标中）是表征病毒释放和CCS所需的pUL 34和pUL 51的功能和相互作用。因此，这些病毒蛋白可用作鉴定也参与的关键病毒和细胞蛋白的工具。第二种方法（包含在第三个具体目标中）是利用最近开发的关于将细胞膜蛋白递送到细胞基底外侧和连接表面的宿主途径的信息，并使用关键分子的显性负抑制剂来探测这些途径。