Cell Type Specific Outcomes of Gammaherpesvirus Infection.

描述（由申请人提供）：潜伏性γ疱疹病毒（？HV）感染存在于>90%的世界人口中，并且与慢性炎症和许多恶性肿瘤相关。关于原发性感染的细胞结果的知识存在显著差距，这限制了预防潜伏感染的治疗方法的发展。我们假设初次手术的结果？HV感染取决于感染的细胞类型。这里概述的实验将提高知识的早期事件？HV感染，使用鼠γ疱疹病毒68（？HV 68）作为一个模型。我们将把我们的调查重点放在B细胞和内皮细胞（EC），因为他们都有作用，在慢性？HV感染和转化的细胞类型很多吗？HV相关肿瘤。我们的初步数据表明yHV 68感染的结果在B细胞和EC中是非常不同的，并且这两种感染都不符合目前的裂解复制与潜伏感染的工作模型。本研究的目的一是探讨yHV 68感染B细胞的早期转归。分析体外和体内感染，我们将利用标记的病毒来确定yHV 68基因组定位于受感染的B细胞核的效率。我们还将通过荧光素酶报告基因和PCR分析确定yHV 68基因组在感染的B细胞中转录的稳健性。本项目的目的2是确定自噬对体外EC感染结果的贡献。我们将确定yHV 68感染的EC幸存者是否正在经历自噬，以及抑制自噬对感染结果的影响。我们还将测试候选自噬相关病毒基因在EC感染结果中的作用。本项目的目的3是表征yHV 68感染体内的EC结果。我们将使用免疫组织化学、原位杂交和RT-PCR检测和表征感染小鼠组织中感染的EC。相关性：慢性yHV感染存在于>90%的人群中，并与许多不同癌症的发生相关。目前，我们对yHV感染的早期结果了解不够，无法设计用于降低慢性疾病相关感染患病率的药物。这些实验很重要，因为它们将确定yHV感染相关特定细胞类型的早期结果。

DESCRIPTION (provided by applicant): Latent gammaherpesvirus (?HV) infection exists in >90% of the world's population, and is associated with chronic inflammation and numerous malignancies. There is a significant gap in knowledge regarding cellular outcomes of primary infection, and this limits the development of therapeutics to prevent latent infection. We hypothesize that outcome of primary ?HV infection is dependent on the cell type infected. The experiments outlined here will enhance knowledge of the early events in ?HV infection, using murine gammaherpesvirus 68 (?HV68) as a model. We will focus our investigation on B cells and endothelial cells (ECs) because they both have roles in chronic ?HV infection and are the transformed cell types of many ?HV-associated tumors. Our preliminary data indicate that outcome of yHV68 infection is very different in B cell and ECs, and that neither infection adheres to the current working model of lytic replication versus latent infection. Aim 1 of this project is to investigate the early outcome of yHV68 infection in B cells. Analyzing both in vitro and in vivo infection, we will make use of labeled virus to determine how efficiently yHV68 genome localizes to the nucleus of infected B cells. We also will determine how robustly yHV68 genome is transcribed in infected B cells via a luciferase reporter and PCR analysis. Aim 2 of this project is to determine the contribution of autophagy to in vitro EC infection outcome. We will determine if EC survivors of yHV68 infection are undergoing autophagy, and what effect inhibiting autophagy has on infection outcome. We also will test candidate autophagy-associated viral genes for their role in EC infection outcome. Aim 3 of this project is to characterize EC outcome of yHV68 infection in vivo. We will detect and characterize infected ECs in tissues from infected mice using immunohistochemistry, in situ hybridization, and RT-PCR. Relevance: Chronic yHV infection exists in >90% of the population and is associated with the development of many different cancers. Currently we do not know enough about the early outcomes of yHV infection to design drugs for reducing the prevalence of chronic, disease-associated infection. These experiments are important because they will determine early outcomes of yHV infection in relevant specific cell types.