ROLE OF B CELLS IN MURINE GAMMAHERPES -68 LATENCY

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。γ疱疹病毒与淋巴细胞增生性疾病和淋巴瘤以及其他癌症的发生密切相关。这项研究的长期目标是了解伽玛疱疹病毒如何操纵正常的B细胞或T细胞发育，从而在受感染宿主的淋巴细胞室中持续存在。了解伽玛疱疹病毒在被感染宿主中持续存在的机制可能会导致干扰慢性感染的策略的发展。对小鼠γ疱疹病毒68的拟议研究的重点是正在努力开发一种可处理的小动物模型，以表征γ疱疹病毒感染的建立和维持。目的1。体内B细胞γ - hv68潜伏期分析。1一个。定量腹腔和鼻内感染后脾脏B细胞和非B细胞潜伏期随时间的变化；和1 b。描述潜伏感染B细胞的表型；目标2。体内γ - hv68潜伏期的定量转换和动态。2 a。生成和表征复制缺陷γ - hv68突变体，以确定体内潜伏库的半衰期；2 b。通过将未感染和潜伏感染的免疫活性小鼠脾细胞过继移植到rag1缺陷小鼠体内分析B细胞潜伏期的传播和2 c。利用基因标记的病毒监测病毒通过B细胞的运输。目标3。评估长期γ - hv68感染对B细胞潜伏期的要求。3 a。生成含有白喉毒素基因的重组病毒；3 b。条件白喉毒素表达病毒在正常小鼠和组织特异性Cre重组酶表达小鼠中的表达分析和3 c。生成并表征有条件表达Cre重组酶活性的小鼠？表达γ - hv68的条件白喉毒素的应用。

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.