HSV and Host Cell Contributions to Corneal Infection

HSV 和宿主细胞对角膜感染的影响

• 批准号: 6744750
• 负责人: JENNIFER Hart LAVAIL
• 金额: $ 26.51万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6744750
• 关键词: axon; cell cell interaction; cell membrane; cell migration; corneal epithelium; electron microscopy; eye infections; eye injury; gene expression; glycoproteins; green fluorescent proteins; herpes simplex virus 1; host organism interaction; immunocytochemistry; intracellular transport; laboratory mouse; light microscopy; monoclonal antibody; plaque assay; protein transport; relapse /recurrence; virus envelope; virus infection mechanism; virus protein

The ability of a virus to usurp the normal cell biology of a host cell for viral replication and spread depends on both viral gene expression and host cell responses. The key to controlling viral invasion of an epithelium lies in understanding the mechanisms by which virus spreads within and between host cells. We shall focus on mechanisms by which Herpes simplex virus type 1 (HSV) invades and usurps the machinery of corneal epithelial cells. The available information about the regulation of HSV infection and spread in the cornea is limited. However, it is clear that at least three mechanisms are involved. 1) Free virus can enter the cornea from a distance, attach and fuse with a host cell plasma membrane and enter the cell (viral fusion mechanism). 2) A different mechanism is used for transfer of virions from an infected cell to coupled adjacent cells (cell-to-cell spread mechanism). 3) Finally, a less widely recognized mechanism involves the movement of virus in an infected cell as the cell moves from one location to another (translational mechanism). We hypothesize that viral envelope glycoproteins and host cell surface receptors play key roles in the spread of HSV in cornea in the first and second mechanisms, but not in the translational mechanism. We will use two mouse model systems: first, primary infection with virus delivered to the corneal surface injury; and second, a model of recurrent herpetic infection by viral delivery to the basal surface of the intact cornea via infected trigeminal ganglion cell axons. Using genetic, pharmacological and immunocytochemical tools, we shall define the contributions of viral fusion and cell-to-cell spread by their different sensitivity to mutations in the HSV envelope glycoproteins. The role of translocation of virally infected cells will be determined by delivering HSV as a pulse, using Valacyclovir to eliminate secondary cell infection. These in vivo experiments will provide fundamental, cell biological information about the transfer of HSV between squamous epithelial cells in vivo and the response of host cell junctions to injury. In addition, the results will also have significant clinical benefits. A better understanding of the differences between spread of virus after corneal injury and of the spread that results from reactivated HSV delivered to the cornea in human herpetic keratitis will focus attention on whether different strategies are required for treatment of initial as opposed to recurrent infections. Moreover, our results will lead to identification of viral and host proteins that are necessary for viral spread and a rational basis for the design of innovative antiviral drugs.

病毒篡夺宿主细胞的正常细胞生物学对病毒复制和扩散的能力取决于病毒基因表达和宿主细胞反应。 控制上皮的病毒侵袭的关键在于了解病毒在宿主细胞内和之间传播的机制。 我们将重点介绍单纯疱疹病毒1型（HSV）入侵并篡夺角膜上皮细胞机械的机制。 有关HSV感染和角膜中传播的调节的可用信息有限。但是，很明显，至少涉及三种机制。 1）游离病毒可以从远处进入角膜，与宿主细胞质膜连接并融合并进入细胞（病毒融合机制）。 2）使用不同的机制将病毒体从感染细胞转移到偶联的相邻细胞（细胞间扩散机制）。 3）最后，随着细胞从一个位置移动到另一个位置（翻译机制），一种不太认识的机制涉及感染细胞中病毒的运动。 我们假设病毒包膜糖蛋白和宿主细胞表面受体在第一和第二机制中HSV在角膜中的扩散中起关键作用，但在翻译机制中却没有。我们将使用两个小鼠模型系统：首先，将病毒带到角膜表面损伤中；其次，通过感染的三叉神经节细胞轴突将病毒递送到完整角膜的基础表面的复发性疱疹感染模型。 使用遗传学，药理和免疫细胞化学工具，我们将通过对HSV包膜糖蛋白中突变的不同敏感性来定义病毒融合和细胞间传播的贡献。 病毒感染细胞的易位的作用将通过使用Valacyclovir消除继发性细胞感染来确定HSV作为脉冲。这些体内实验将提供有关HSV在体内鳞状上皮细胞之间转移以及宿主细胞连接对损伤的反应的基本细胞生物学信息。 此外，结果还将具有重大的临床益处。 更好地理解角膜损伤后病毒传播与人类疱疹性角膜炎角膜的差异之间的差异将集中注意是否需要不同的策略来治疗初始感染，而不是经常性感染。 此外，我们的结果将导致鉴定病毒蛋白和宿主蛋白，这些蛋白是病毒传播所必需的，并且是设计创新抗病毒药的合理基础。