Role of B cells in murine gammaherpes-68 latency

描述（由申请人提供）： γ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γ疱疹病毒如何操纵正常的B或T细胞发育，使其在受感染宿主的淋巴区室中持续存在。了解γ疱疹病毒在受感染宿主中持续存在的机制可能会导致干预慢性感染的策略的发展。拟定的鼠γ疱疹病毒68（γ HV 68;也称为MHV-68）研究的重点代表了正在努力开发一种易于处理的小动物模型，用于表征γ疱疹病毒感染的建立和维持。 目标1。分析体内B细胞中的γ HV 68潜伏期。1.a.定量脾中B细胞和非B细胞潜伏期作为腹膜内和鼻内感染后时间的函数;和1.B.表征潜伏感染的B细胞的表型;目的2.定量体内γ HV 68潜伏期的转换和动力学; 2. a.产生和表征复制缺陷型oHV 68突变体以确定体内潜伏储库的半衰期; 2. b.通过将脾细胞从幼稚和潜伏感染的免疫活性小鼠过继转移到Rag 1缺陷小鼠中来分析B细胞潜伏期的扩散;和2. c.使用基因标记病毒监测通过B细胞的病毒运输。 目标3。评估长期γ HV 68感染对B细胞潜伏期的要求。3.a.产生携带条件表达的白喉毒素基因的重组病毒; 3.b.分析正常小鼠和表达组织特异性Cre重组酶的小鼠中表达条件性白喉毒素的病毒;和3. c.用表达γ HV 68的条件性白喉毒素产生和表征条件性表达cre重组酶活性的小鼠-利用。

DESCRIPTION (provided by applicant): 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 manipulates 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 (gammaHV68; also referred to as MHV-68) represents an ongoing effort to develop a tractable small animal model for characterizing establishment and maintenance of gammaherpesvirus infection. Aim 1. Analyze gammaHV68 latency in B cells in vivo. 1.a. Quantitate B cell and non-B cell latency in the spleen as a function of time after intraperitoneal and intranasal infection; and 1.b. Characterize the phenotype(s) of latently infected B cells; Aim 2. Quantitate turnover and dynamics of gammaHV68 latency in vivo; 2.a. Generate and characterize replication-deficient oHV68 mutants to determine the half-life of latency reservoirs in vivo; 2.b. Analyze spread of B cell latency by adoptive transfer of splenocytes from naive and latently infected immunocompetent mice into Rag 1-deficient mice; and 2.c. Monitor virus trafficking through B cells using genetically-marked viruses. Aim 3. Assess the requirement for B cell latency for long-term gammaHV68 infection. 3.a. Generate recombinant viruses harboring a conditionally expressed diphtheria toxin gene; 3.b. Analyze conditional diphtheria toxin expressing viruses in normal and tissue-specific Cre recombinase expressing mice; and 3.c. Generate and characterize mice that conditionally express cre recombinase activity - utilization with conditional diphtheria toxin expressing gammaHV68.