Gene Expression in HSV-1 Latency After Corneal Infection

角膜感染后 HSV-1 潜伏期的基因表达

• 批准号: 6861733
• 负责人: Paul R. Kinchington
• 金额: $ 28.97万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-01 至 2008-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6861733
• 关键词: antigen presentation; confocal scanning microscopy; cornea disorder; cytotoxic T lymphocyte; fluorescence microscopy; gene expression; gene induction /repression; green fluorescent proteins; herpes simplex virus 1; in situ hybridization; keratitis; laboratory mouse; latent virus infection; neuroimmunomodulation; protein biosynthesis; recombinant virus; trigeminal nerve; virus genetics; virus infection mechanism; virus protein

DESCRIPTION (provided by applicant): Herpes simplex virus type 1 (HSV-1) is a major cause of corneal blindness. In humans, corneal disease results from repeated reactivation of HSV-1 from a latent (quiescent) state that was established in the host sensory ganglia at the time of primary infection. Recent findings from several laboratories, including ours, challenge the general supposition that a lack of viral protein synthesis during HSV-1 latency precludes immune detection of latently infected neurons and immune modulation of the latent state. Rather, these findings give rise to a dynamic model of latency in which many reactivation attempts are aborted by a preemptive CD8+ T cell response in the latently infected ganglia. Our underlying hypothesis is that gene expression during latency allows ganglionic CD8+T cells to provide a 'just in time' mechanism to inhibit virus production in vivo. In this proposal, the concepts of antigen expression during latency and the enhancement of the immune-mediated maintenance of viral latency will be explored. Specific Aim 1 will test the hypothesis that gene expression occurs in the TG during latency and induced reactivation without the production of infectious virus and 'true late' viral genes. We will derive recombinant HSV-1 that regulate two fluorescent proteins from different candidate viral promoters. Mice will be infected through the corneal route to establish latency. In conjunction with confocal imaging techniques, we will identify HSV-1 promoter activities in the TG of latently infected and reactivation-induced mice. Our model predicts that specific HSV-1 promoters are active during murine latency in the absence of late (gamma-2) genes and infectious virus. In Specific Aim 2, we will test the hypothesis that immune infiltration of CD8+ T cells into the ganglia, and the protection they afford from reactivation, can be enhanced by persistent expression of viral epitopes in latently infected neurons. Viruses will be developed which express multimers of an immunodominant peptide under latency active promoters. Mice latently infected with such viruses will be assessed for CD8+ T cell infiltration and resistance to induced reactivation. Such studies will establish foundations for the design of vaccines to augment immune regulation of latency and reactivation.

描述（由申请人提供）：单纯疱疹病毒1型（HSV-1）是角膜失明的主要原因。在人类中，角膜疾病是由于原发性感染时在宿主感觉神经节中建立的潜在（静止）状态的HSV-1重新激活而引起的。来自几个实验室的最新发现，包括我们的实验室，挑战了总体上的假设，即HSV-1潜伏期期间缺乏病毒蛋白合成可阻止对潜在感染神经元的免疫检测和潜在状态的免疫调节。相反，这些发现产生了一种动态的潜伏模型，其中许多重新激活的尝试被潜在感染的神经节中的先发性CD8+ T细胞反应中止。我们的基本假设是，潜伏期期间的基因表达允许神经节CD8+T细胞提供一种“及时”的机制来抑制体内病毒的产生。在此提案中，将探索潜伏期期间抗原表达的概念以及免疫介导的病毒潜伏期维持的增强。具体目标1将检验以下假设：潜伏期期间TG发生基因表达并引起重新激活，而没有产生传染病病毒和“真实的晚期”病毒基因。我们将得出重组HSV-1，该HSV-1调节来自不同候选病毒启动子的两种荧光蛋白。小鼠将通过角膜路线感染以建立潜伏期。结合共聚焦成像技术，我们将确定在潜在受感染和重新激活诱导的小鼠Tg中的HSV-1启动子活性。我们的模型预测，在没有近来（伽马2）基因和传染性病毒的情况下，特定的HSV-1启动子在鼠潜伏期期间具有活性。在特定的目标2中，我们将检验以下假设：CD8+ T细胞免疫浸润到神经节中，并且可以通过在潜在感染的神经元中持续表达病毒表位来增强其免受重新激活的保护。将开发病毒，在潜伏期活跃启动子下表达免疫主导肽的多聚体。将评估受到此类病毒感染的小鼠的CD8+ T细胞浸润和对诱导重生的耐药性。此类研究将为设计疫苗的设计基础，以增强潜伏和重新激活的免疫调节。