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: 6719312
• 负责人: Paul R. Kinchington
• 金额: $ 29.03万
• 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/6719312
• 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）：1 型单纯疱疹病毒 (HSV-1) 是导致角膜失明的主要原因。在人类中，角膜疾病是由于 HSV-1 从初次感染时在宿主感觉神经节中建立的潜伏（静止）状态反复重新激活所致。包括我们在内的多个实验室的最新研究结果对以下普遍假设提出了挑战：HSV-1 潜伏期病毒蛋白合成的缺乏阻碍了对潜伏感染神经元的免疫检测和潜伏状态的免疫调节。相反，这些发现产生了一个动态潜伏模型，其中许多重新激活尝试都被潜伏感染神经节中先发制人的 CD8+ T 细胞反应所中止。我们的基本假设是，潜伏期的基因表达允许神经节 CD8+T 细胞提供“及时”机制来抑制体内病毒的产生。在该提案中，将探讨潜伏期抗原表达的概念以及增强免疫介导的病毒潜伏期维持。具体目标 1 将检验以下假设：基因表达发生在潜伏期间的 TG 中，并诱导重新激活，而不会产生传染性病毒和“真正的晚期”病毒基因。我们将从不同的候选病毒启动子中衍生出调节两种荧光蛋白的重组HSV-1。小鼠将通过角膜途径被感染以建立潜伏期。结合共聚焦成像技术，我们将鉴定潜伏感染和重新激活小鼠的 TG 中的 HSV-1 启动子活性。我们的模型预测，在缺乏晚期 (gamma-2) 基因和传染性病毒的情况下，特定的 HSV-1 启动子在小鼠潜伏期是活跃的。在具体目标 2 中，我们将测试以下假设：CD8+ T 细胞免疫浸润到神经节中，以及它们提供的免于重新激活的保护，可以通过病毒表位在潜伏感染神经元中的持续表达来增强。将开发在潜伏活性启动子下表达免疫显性肽多聚体的病毒。将评估潜伏感染此类病毒的小鼠的 CD8+ T 细胞浸润和对诱导再激活的抵抗力。此类研究将为疫苗设计奠定基础，以增强潜伏期和再激活的免疫调节。