ROLE OF B CELLS IN MURINE GAMMAHERPES -68 LATENCY

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。伽马疱疹病毒与淋巴增生性疾病和淋巴瘤以及其他癌症的发展密切相关。这项研究的长期目标是了解伽马疱疹病毒如何操纵正常的B或T细胞发育，使其持续存在于受感染宿主的淋巴间隔内。了解伽玛疱疹病毒在受感染宿主中持续存在的机制可能会导致制定干预慢性感染的策略。拟议的关于小鼠伽马疱疹病毒68的研究的重点是正在进行的努力，以开发一种易驯化的小动物模型，以表征伽马疱疹病毒感染的建立和维持。目的1.分析活体B细胞中γ-HV68的潜伏期。1A.量化B细胞和非B细胞在脾内和鼻腔感染后潜伏期随时间的变化；研究潜伏感染B细胞的表型特征(S)；目的2.定量检测γ-HV68潜伏期在体内的周转和动态。2A。产生并鉴定复制缺陷的伽马-HV68突变体，以确定体内潜伏库的半衰期；2b.分析通过过继转移初治和潜伏感染免疫活性小鼠的脾细胞到RAg 1缺陷小鼠的B细胞潜伏时间的传播；以及2c.使用基因标记的病毒监测通过B细胞的病毒传播。目的3.评估长期感染Gamma-HV68对B细胞潜伏期的要求。3A.产生携带有条件表达的白喉毒素基因的重组病毒；在正常和组织特异性Cre重组酶表达的小鼠中分析条件白喉毒素表达病毒；以及3c.产生并鉴定有条件表达Cre重组酶活性的小鼠？利用条件性白喉毒素表达伽马-hv68。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Gammaherpesviruses are closely associated with the development of lymphoproliferative disease and lymphomas, as well as other cancers. The long-term goal of this research is to understand how gammaherpesviruses manipulate normal B or T cell development to persist within the lymphoid compartment of the infected host. Understanding the mechanisms used by gammaherpesviruses to persist in the infected host may lead to the development of strategies for interfering with chronic infection. The focus of the proposed studies on murine gammaherpesvirus 68 represents an ongoing effort to develop a tractable small animal model for characterizing establishment and maintenance of gammaherpesvirus infection. Aim 1. Analyze gamma-HV68 latency in B cells in vivo. 1a. Quantitate B cell and non-B cell latency in the spleen as a function of time after intraperitoneal and intranasal infection; and 1b. Characterize the phenotype(s) of latently infected B cells; Aim 2. Quantitate turnover and dynamics of gamma-HV68 latency in vivo. 2a. Generate and characterize replication-deficient gamma-HV68 mutants to determine the half-life of latency reservoirs in vivo; 2b. Analyze spread of B cell latency by adoptive transfer of splenocytes from na¿ve and latently infected immunocompetent mice into Rag 1-deficient mice; and 2c. Monitor virus trafficking through B cells using genetically-marked viruses. Aim 3. Assess the requirement for B cell latency for long-term gamma-HV68 infection. 3a. Generate recombinant viruses harboring a conditionally expressed diphtheria toxin gene; 3b. Analyze conditional diphtheria toxin expressing viruses in normal and tissue-specific Cre recombinase expressing mice; and 3c. Generate and characterize mice that conditionally express Cre recombinase activity ? utilization with conditional diphtheria toxin expressing gamma-HV68.