INTERCELLULAR TRANSPORT OF HSV-1 IN CORNEAL EPITHELIUM

这项建议的目的是确定HSV-1在角膜上皮反复疱疹感染过程中细胞间传播的机制。反复发生的角膜疱疹感染是导致人类失明的主要原因。了解病毒在角膜中细胞间传播的机制对预防这种疾病具有重要意义。第一个具体目标是确定角膜上皮细胞中被病毒用于细胞间传播的因素，例如与细胞连接相关的蛋白质。第二个目标是针对涉及细胞间传播的病毒基因产品。第三个目标是确定感染的角膜细胞在细胞间传播过程中的形态变化。最后一个目标是开发和实施一种病毒感染的培养系统，模拟眼睛的自然条件，以研究病毒从受感染的感觉神经元轴突向角膜上皮细胞的传播。人角膜上皮细胞在培养过程中将接种单纯疱疹病毒类型。1.将使用与细胞连接相关的宿主蛋白的特定抑制剂，并确定它们对病毒在细胞间传播的影响。疱疹病毒突变株将用于阐明病毒传播所必需的特定病毒基因产品。被感染的角膜细胞的形态变化将通过光学、共聚焦和电子显微镜进行监测。最后，将开发一种新的培养系统，模拟体内反复发生的角膜疱疹感染。了解病毒是如何在眼睛内细胞间传播的，以及涉及的特定宿主和病毒基因，对于开发有效的治疗复发性角膜疱疹感染具有重要意义。

The goal of this proposal is to define the mechanisms involved in cell-to-cell transmission of HSV-1 during recurrent herpetic infection of the corneal epithelium. Recurrent herpetic infection of the cornea is a leading cause of blindness in humans. Understanding the mechanisms of cell-to-cell spread of virus in the cornea will be important in preventing this disease. The first specific aim is to identify factors in corneal epithelial cells that are utilized by the virus for cell-to-cell spread, such as proteins associated with cell junctions. The second aim is to target viral gene products involved cell-to-cell spread. The third aim focuses on identifying morphologic changes in infected corneal cells during cell-to-cell spread. The last aim is to develop and implement a culture system of viral infection that models the natural conditions of the eye to study the spread of virus from infected sensory neuron axons to corneal epithelial cells. Human corneal epithelial cells in culture will be inoculated with herpes simplex virus type. 1. Specific inhibitors of host proteins associated with cell junctions will be administered and their effects on cell-to-cell spread of virus determined. Mutant strains of herpes virus will be used to clarify specific viral gene products essential for viral spread. Morphological changes in the infected corneal cells will be monitored by light, confocal and electron microscopy. Finally, a novel culture system that models recurrent herpetic infection of cornea in vivo will be developed. Understanding how virus is spread cell-to-cell in the eye and the specific host and viral genes involved will be important for the development of effective treatments for recurrent corneal herpetic infection.