HSV INDUCED RNA DEGRADATION AND PATHOGENESIS

HSV 诱导的 RNA 降解和发病机制

• 批准号: 2711121
• 负责人: David A Leib
• 金额: $ 22.4万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2711121
• 关键词: confocal scanning microscopy; gene deletion mutation; herpes simplex virus 1; herpes simplex virus 2; host organism interaction; immunocytochemistry; keratitis; laboratory mouse; latent virus infection; mutant; neurotropic virus; ocular herpes; open reading frames; recombinant virus; site directed mutagenesis; tissue /cell culture; virion; virus RNA; virus cytopathogenic effect; virus genetics; virus protein

DESCRIPTION (from abstract): Herpes simplex virus (HSV) keratitis is a leading cause of non-traumatic blindness in the US, with more than 200,000 cases per year. HSV can cause a variety of ocular diseases in humans ranging from self-limiting dendritic epithelial keratitis, conjunctivitis, and blepharitis to necrotizing stromal keratitis. In addition, HSV commonly causes cold sores, genital sores, and is a leading cause of viral encephalitis. The life cycles of HSV and other neurotropic herpesviruses are characterized by a lytic phase of infection at peripheral sites such as the cornea and skin during which all virus genes are expressed, and a latent phase of infection in neurons, during which gene expression is extremely limited. Latency represents a lifelong source of virus which can reactivate periodically causing severe ocular and other mucocutaneous damage, and the ability to establish lifelong latency renders HSV resistant to cure. One hallmark of the neurotropic herpesviruses is their ability to rapidly shut off macromolecular synthesis in the cells that they infect. It has been shown for HSV type 1 (HSV-1) that the gene responsible for this shut off is the product of the UL41 gene known as the virion host shutoff protein or vhs. Vhs is an important determinant of pathogenicity, allowing the virus to replicate to high titers in the cornea in vivo, to damage the cornea, and to establish latency with high efficiency. The objectives of this proposal are to investigate further the mechanisms of action and functions of the vhs protein in vivo. To this end, mutations will be introduced into the open reading frame of vhs as well as into other genes which are required for and associated with vhs activity. In addition, intertypic and interstrain recombinants will be generated in which vhs is exchanged from one virus to another. The effect of these mutations and exchanges upon the ability of the virus to induce degradation of host and viral mRNAs will then be measured by in vitro express/degradation assays and in vivo following introduction of these mutations into the context of the viral genome. Resulting mutants will also be tested for their ability to establish, maintain, and reactivate from viral latency in neurons. Efforts will also be made to assess the role of vhs in the induction of corneal damage and the role of interferons in limiting corneal disease. A better understanding of vhs will allow further insight into the mechanisms by which HSV can persist for the lifetime of its host and indicate novel therapeutic approaches and targets for control of this blinding disease.

描述（来自摘要）：单纯疱疹病毒（HSV）角膜炎是一种 是美国非创伤性失明的主要原因， 案件每年。 单纯疱疹病毒可引起人类多种眼部疾病 从自限性树突状上皮角膜炎，结膜炎， 以及睑缘炎到坏死性角膜基质炎。 此外，HSV通常 导致唇疱疹，生殖器溃疡，是病毒性疾病的主要原因， 脑炎 HSV和其他嗜神经性疱疹病毒的生命周期 其特征在于在外周部位感染的溶解期， 角膜和皮肤，在此期间，所有病毒基因都被表达， 在神经元中的感染阶段，在此期间，基因表达是非常重要的。 有限公司 潜伏期代表了病毒的终身来源， 周期性地引起严重的眼部和其他皮肤粘膜损伤， 建立终身潜伏期的能力使HSV对治愈具有抗性。 嗜神经性疱疹病毒的一个标志是它们能够快速地 关闭被感染细胞中的大分子合成。 它有 对于HSV 1型（HSV-1），已经表明负责这种关闭的基因 关闭是UL 41基因的产物，称为病毒体宿主关闭蛋白 或VHS。 VHS是致病性的重要决定因素， 病毒在体内角膜中复制到高滴度，破坏角膜， 角膜，并建立高效率的潜伏期。 本提案的目的是进一步研究 VHS蛋白在体内的作用和功能。 为此，突变 将被引入到VHS的开放阅读框架中以及其它 VHS活性所需的和与VHS活性相关的基因。 此外，本发明还提供了一种方法， 将产生型间和株间重组体， 从一种病毒到另一种病毒。 这些突变的影响， 病毒诱导宿主降解的能力， 然后通过体外表达/降解测定来测量病毒mRNA， 在将这些突变引入到本发明的背景中之后， 病毒基因组 产生的突变体还将接受测试，以确定它们是否有能力 在神经元中建立、维持并从病毒潜伏期中重新激活。 努力 还将评估VHS在诱导角膜炎中的作用。 损伤和干扰素在限制角膜疾病中的作用。 更好的 对VHS的理解将使我们能够进一步了解 HSV可在其宿主的一生中持续存在，并指示新的治疗方法。 控制这种致盲性疾病的方法和目标。