Cell Type Specific Outcomes of Gammaherpesvirus Infection.

描述（由申请人提供）：潜伏γ疱疹病毒(？乙型肝炎病毒（HV）感染存在于世界90%以上的人口中，并与慢性炎症和许多恶性肿瘤有关。关于原发性感染的细胞结果的知识有很大的差距，这限制了治疗方法的发展，以防止潜伏性感染。我们假设primary ？hiv感染取决于被感染的细胞类型。这里概述的实验将增强对早期事件的认识。小鼠γ疱疹病毒68 （?）HV68)作为模型。我们将重点研究B细胞和内皮细胞（ECs），因为它们都在慢性？hiv感染的转化细胞类型有很多吗？HV-associated肿瘤。我们的初步数据表明，yHV68感染在B细胞和ec中的结果非常不同，并且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。该项目的目的1是研究B细胞感染yHV68的早期结果。分析体外和体内感染，我们将利用标记病毒来确定yHV68基因组定位到感染B细胞细胞核的效率。我们还将通过荧光素酶报告基因和PCR分析来确定yHV68基因组在感染B细胞中的转录强度。该项目的目的2是确定自噬对体外EC感染结果的贡献。我们将确定感染yHV68的EC幸存者是否正在进行自噬，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。该项目的目的3是表征体内感染yHV68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的感染内皮细胞。相关性：慢性yHV感染存在于50%至90%的人群中，并与许多不同癌症的发展有关。目前，我们对yHV感染的早期结果知之甚少，无法设计出降低慢性疾病相关感染流行率的药物。这些实验很重要，因为它们将确定相关特定细胞类型中yHV感染的早期结果。

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.