Transport of Herpes Simplex Virus in Ocular Tissues

描述（由申请人提供）： 了解嗜神经病毒的轴突运输机制是 这是理解和控制它们在神经系统中传播的关键。到 繁殖和传播，最初从入口处移动，后来移动到 单纯疱疹病毒（HSV）释放位点必须利用神经元宿主细胞 蛋白质和机制。我们对特定病毒相互作用的理解 这些蛋白质与神经元蛋白质的机制有关， 被剥削，是至关重要的，但几乎不存在。在本申请中，我们继续 专注于HSV的顺行运输。根据我们之前的结果， 我们的假设是HSV的核衣壳和包膜成分是 在轴突中独立运输，并且组件需要特定的 驱动蛋白相关蛋白1)我们已经开始检查 核衣壳和包膜成分。为此，我们开发了两种病毒 突变株和回复突变体，这将有助于我们对这些 运输机制。2)在本建议中，我们将继续进行这些研究， 进行免疫共沉淀试验以鉴定马达蛋白 与核衣壳成分相关。3)我们还将确定 或者病毒不是通过出芽从细胞中排出， 被传递到轴突膜，核衣壳聚集在轴突膜附近， 膜的区域。 这些结果将提供重要的新的细胞生物学信息， 特定细胞器的识别信号。他们也将有重大的 临床获益。单纯疱疹病毒在人角膜中的顺行转运 疱疹性角膜炎导致严重的后果，包括角膜瘢痕， 青光眼可能还有脑炎我们的研究结果将提供新的见解， 鉴定病毒包膜所必需的病毒和宿主蛋白， 核衣壳运输和创新设计的合理依据 抗病毒药物的预防和干预。此外，HSV的基因组 可以被改变以用作将新基因引入到细胞中的载体。 神经系统我们的研究结果将阐明靶向的机制， 特定的神经元类型和感染的特定区域 神经元

DESCRIPTION (provided by applicant): Understanding the mechanisms of axonal transport of neurotropic viruses is the key to understanding and controlling their spread in the nervous system. To multiply and spread, to move initially from the site of entry and later to the site of release, Herpes simplex virus (HSV) must use neuronal host cell proteins and mechanisms. Our understanding of the interplay of specific viral proteins that piggyback on the mechanisms and of the neuronal proteins that are exploited, is critical but almost nonexistent. In this application we continue to focus on the anterograde transport of HSV. Based on our previous results, our hypothesis is that the nucleocapsid and envelope components of HSV are independently transported in the axon and that the components require specific kinesin related proteins. 1) We have begun to examine the transport of the nucleocapsid and envelope components. To do this we have developed two viral mutant strains and revertants that will facilitate our research into these transport mechanisms. 2) In this proposal we shall continue these studies and carry out co-immunoprecipitation assays to identify the motor proteins associated with the nucleocapsid component. 3) We shall also determine whether or not the virus egresses from the cell by budding, after the envelope proteins are delivered to the axon membrane and the nucleocapsids cluster near that region of membrane. These results will provide important new cell biological information about the recognition signals of particular organelles. They will also have significant clinical benefits. The anterograde transport of HSV to the cornea in human herpetic keratitis results in severe consequences, including corneal scars, glaucoma and possibly encephalitis. Our results will provide new insight into the identification of viral and host proteins necessary for viral envelope and nucleocapsid transport and a rational basis for the design of innovative antiviral drugs for prevention and intervention. Furthermore, the genome of HSV can be altered to serve as a vector for introduction of novel genes into the nervous system. Our results will elucidate the mechanisms that target the vector to particular neuron types and to particular regions of infected neurons.