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（由申请人提供）：αHerpeSvirus病的所有表现（单纯疱疹，绒毛皮带轮）是由于病毒从粘膜表面的初始感染或细胞传播到该区域中其他细胞的能力，并使神经细胞神经细胞的神经细胞蔓延。症状的复发和随之而来的病毒传播给新宿主类似，需要能够从感觉神经节细胞传播到外围的细胞以及粘膜表面上的细胞中。令人惊讶的是，该病毒在复发感染中的扩散发生在适应性免疫反应时，包括应中和从细胞中释放的病毒的抗体反应。因此，这些病毒的引起疾病的特性取决于用于保护病毒免受适应性免疫反应效应子的细胞传播的机制。 人α在宿主中的传播需要从高尔基体的组装部位运输新组装的病毒颗粒，以暴露于细胞表面，以释放到细胞外培养基或细胞连接处以进行细胞间传播（CCS）。这两种贩运途径都被众所周知。在 部分是因为，除了将病毒进入宿主细胞所需的病毒蛋白外，在大多数细胞类型中未发现该过程所需的VIRL基因功能。我们发现，两个病毒基因产物PUL34和PUL51在有效的病毒释放和/或CC中起关键作用。两种蛋白质显然都是多功能的。 PUL34是疱疹病毒衣壳的核出口所必需的，并且已证明PUL51是有效的病毒细胞质组装所必需的。但是，我们发现两种蛋白质在释放和CC中都起着至关重要的作用，这些蛋白质可以与其在病毒体组装中的作用上遗传上偶联。 我们的总体目标是检验以下假设：HSV释放和CC是通过对细胞途径的病毒劫持来实现的，这些途径将宿主细胞膜蛋白分类为适当的表面和连接。我们将使用两种一般方法来实现这一总体目标。第一种方法（包含在前两个特定目的中）是表征病毒释放和CC所需的PUL34和PUL51的功能和相互作用。因此，这些病毒蛋白可以用作识别也参与的关键病毒和细胞蛋白的工具。第二种方法（第三个特定目的中包含的方法是利用有关宿主途径的最新信息，这些信息将细胞膜蛋白传递到细胞的基底外侧和连接式表面，并使用关键分子的主要负抑制剂探测这些途径。