HERPES SIMPLEX VIRUS AND NGF DIFFERENTIATED PC12 CELL INTERACTION

单纯疱疹病毒和 NGF 分化的 PC12 细胞相互作用

• 批准号: 6651791
• 负责人: Timothy M Block
• 金额: $ 20.08万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6651791
• 关键词: Herpes simplex disease; PC12 cells; cell age; cell differentiation; circular DNA; gel electrophoresis; gene expression; genome; herpes simplex virus 1; laboratory mouse; latent virus infection; mutant; nerve growth factors; neurotropic virus; polymerase chain reaction; transcription factor; trigeminal nerve; virus DNA; virus genetics; virus infection mechanism; western blottings

Description (provided by applicant) This project is concerned with how neuronal cells manage HSV infection at the intracellular level. The intracellular details of HSV/neuronal cell interactions are difficult to study in vivo. Information regarding the mechanisms and kinetics of HSV genome physical organization following neuronal cell infection in vivo and even in vitro is extremely limited. We have, therefore, developed a tissue culture system of quiescent HSV infection using nerve growth factor (NGF) differentiated cells. This system will be used to determine the impact of HSV upon NGF differentiated cells and to track the fate and structure of viral DNA following infection. Briefly, NGF differentiated PC 12 cells have been shown to support long-term "quiescent" infections of HSV-1. NGF differentiated PC12 cells are not killed by virus infection and, surprisingly, persist longer than uninfected controls. There is little viral transcription and progeny is not detected in the culture medium, despite the presence of an inducible infectious genome. Strangely, the viral genome in quiescently infected PC12 cells persists as a linear form for several weeks before ultimately assuming an endless, presumably circular, state. These cells will, therefore, be used to study (a) if and how HSV can cause populations of PC12 cells to have a survival advantage over uninfected populations; (b) how linear viral genomes can be maintained intact, for weeks in neuronal like cells; (c) the mechanism(s) involved in their assumption of an endless, possibly modified, quiescent viral genomic state. Observations made in this in vitro system will be related to in vivo mouse models of latency by comparing the physical properties of viral DNA derived from tissue derived from infected mice with that from quiescently infected PC12 cells. This work will thus allow for the testing of hypotheses made about HSV latency seen in the vitro system, in mouse models of latency. Some of the information being uncovered in the in vitro, quiescent infection system has the exciting potential to influence our understanding of how HSV genomes are organized and "silenced".

说明（申请人提供） 这个项目关注的是神经元细胞如何管理HSV感染， 细胞内水平。HSV/神经元细胞的细胞内细节 相互作用很难在体内研究。性的信息 HSV基因组物理组织的机制和动力学 体内甚至体外的细胞感染是极其有限的。我们有， 因此，开发了一种静止期HSV感染的组织培养系统， 神经生长因子（NGF）分化细胞。该系统将用于 确定HSV对NGF分化的细胞的影响，并追踪其命运。 和病毒DNA的结构。简而言之，NGF分化PC 12细胞已显示支持HSV-1的长期“静止”感染。 NGF分化的PC 12细胞不被病毒感染杀死， 令人惊讶的是，比未感染的对照组持续时间更长。几乎没有病毒 在培养基中没有检测到转录和后代，尽管 存在可诱导的感染性基因组。奇怪的是， 静止感染的PC 12细胞以线性形式持续数周 最终呈现出一种无止境的、大概是循环的状态。这些细胞 因此，将用于研究（a）HSV是否以及如何引起 PC 12细胞比未感染群体具有存活优势;（B）如何 线性病毒基因组可以在神经元样细胞中保持完整数周， 细胞;（c）参与其假设的机制， 可能是经过修饰的静止病毒基因组状态。在这方面所作的观察 将体外系统与体内小鼠潜伏期模型进行比较 病毒DNA的物理性质来自感染的组织， 静止感染PC 12细胞的小鼠。这项工作将使 为了检验关于在体外系统中观察到的HSV潜伏期的假设， 在小鼠潜伏期模型中。一些信息被发现在 体外，静止感染系统具有令人兴奋的潜力， 了解HSV基因组如何组织和“沉默”。