HERPES SIMPLEX VIRUS ENTRY VIA ENDOCYTOSIS

描述（由申请人提供）：单纯疱疹病毒（HSV）仍然是世界范围内引起人类疾病的主要人类病原体，包括唇疱疹、眼部和生殖器感染、新生儿感染和脑炎。HSV建立终身潜伏感染，其周期性地重新激活以产生复发性疾病。本研究的长期目标是确定HSV进入宿主细胞的机制。了解HSV感染的初始事件可能有助于开发新的治疗方法，并有助于基因治疗方案的病毒靶向方法。HSV长期以来被认为通过与质膜的pH中性融合进入一些细胞。最近，我们发现一些常见的培养细胞类型的初始感染需要内吞作用和低细胞内pH值。因此，HSV使用一种以上的途径将其基因组递送到细胞内部。HSV感染许多不同的培养细胞类型，并且在体内具有多个靶标，包括上皮和神经元来源的细胞。这种相对广泛的宿主范围可以用多种进入途径来解释。HSV通过内吞作用进入的过程包括两个连续的阶段：完整的包膜病毒从细胞表面内化，以及衣壳渗透到胞质溶胶中。我认为这两个阶段是可分离的，并且具有不同的分子要求。拟议研究的具体目的是：（1）表征HSV进入人上皮细胞的细胞机制;（2）确定HSV通过内吞作用进入的病毒要求。将使用分子、生物化学和细胞生物学方法的组合来实现这些目标。结果将定义单纯疱疹病毒进入相关人类细胞类型的决定因素。此外，这些研究应该阐明将传入病毒靶向适当的细胞区室以启动生产性感染的机制。

DESCRIPTION (provided by applicant): Herpes simplex virus (HSV) remains a major human pathogen world-wide causing human diseases including cold sores, ocular and genital infections, neonatal infections and encephalitis. HSV establishes a life-long latent infection which reactivates periodically to produce recurrent disease. The long-term objective of this research is to define the mechanism of HSV entry into host cells. Understanding of the initial events in HSV infection may help develop new treatments and aid with viral targeting approaches for gene therapy protocols. HSV has long been recognized to enter some cells by pH-neutral fusion with the plasma membrane. Recently, we found that initial infection of some common cultured cell types requires endocytosis and low intracellular pH. Thus, HSV uses more than one pathway to deliver its genome to the cell interior. HSV infects many diverse cultured cell types and has multiple targets in vivo including cells of epithelial and neuronal origin. This relatively broad host range may be explained by multiple entry pathways. The process of HSV entry via endocytosis includes two sequential stages: internalization of intact, enveloped virus from the cell surface, and penetration of the capsid into the cytosol. I propose that these two stages are separable and have distinct molecular requirements. The specific aims of the proposed research are: (1) to characterize the cellular mechanism of HSV entry into human epithelial cells; and (2) to identify the viral requirements of HSV entry via endocytosis. A combination of molecular, biochemical and cell biological approaches will be used to achieve these goals. The results will define the determinants for entry of HSV into relevant human cell types. Additionally, these studies should elucidate the mechanisms that target the incoming virus to the proper cellular compartments to initiate productive infection.