Murine Model of Gammaherpesvirus Latency

伽马疱疹病毒潜伏期小鼠模型

• 批准号: 6514926
• 负责人: JEFFERY T SAMPLE
• 金额: $ 26.19万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6514926
• 关键词: B lymphocyte; Burkitt's lymphoma; Epstein Barr virus; Kaposi's sarcoma; dendritic cells; disease /disorder model; gene induction /repression; host organism interaction; human herpesvirus 8; laboratory mouse; latent virus infection; macrophage; microorganism immunology; polymerase chain reaction; recombinant virus; southern blotting; virus genetics

DESCRIPTION (provided by applicant): The long-term objective of the proposed research is to define the mechanisms that enable the gamma herpesviruses to persist indefinitely within their immunocompetent hosts. These mechanisms underlie the pathogenesis and oncogenic potential associated with these viruses, particularly in individuals with acquired or inherited immunodeficiencies. Unfortunately, due to the extreme host restrictions of the human gamma-herpesviruses Epstein-Barr virus (EBV) and Kaposi's sarcoma-associated herpesvirus (KSHV), it has not been possible to directly address the mechanisms responsible for EBV and KSHV persistence in a relevant host setting. Infection of laboratory mice with the murine gamma-herpesvirus MHV-68 (closely related to KSHV) offers a highly tractable and potential model of human gamma-herpesvirus persistence, but viral latency in this system has not been sufficiently characterized at the molecular level to enable exploitation of this model for these purposes. Therefore, the immediate goals of the proposed research are to define the molecular characteristics of MHV-68 latency. To obtain our objective, we propose three specific aims. Under Aim 1 we will define the programs of MHV-68 latency gene expression during the establishment and maintenance phases of latency within the hematopoietic-cell reservoirs of latent virus (activated and resting B cells, macrophages, and dendritic cells). Specifically, we will screen these cells by RT-PCR and Southern blot hybridization for expression of the MHV-68 latency genes M2, M3, M1 1, 72, 73 and 74. Additionally, we propose to identify potentially novel MHV-68 latency genes expressed in these cells that are not apparent from analysis of the viral genornic DNA sequence. Under Aim 2 we will address the functions of three latency-associated genes: M2, 73 and 74, for which little or no information is available. Under Aim 3 we will define the impact of MHV-68 latency on cellular gene expression in vivo at the cellular level. Together, these specific aims will fill the significant gaps that currently remain in our knowledge of the latent life cycle of MHV-68, as well as provide better insight into the mechanisms that the gamma-herpesviruses as a group exploit to maintain latency, and thus contribute to their pathogenic potential.

描述（由申请人提供）：拟议的长期目标 研究是定义使伽马疱疹病毒达到的机制 在其免疫能力的宿主中无限期地持续存在。这些机制 与这些相关的发病机理和致癌潜力的基础 病毒，特别是在被收购或继承的人 免疫缺陷。不幸的是，由于极端的主机限制 人γ-疱疹病毒爱泼斯坦 - 巴尔病毒（EBV）和卡波西 与肉瘤相关的疱疹病毒（KSHV），不可能直接 解决导致EBV和KSHV持久性的机制 主机设置。用鼠γ-鞘膜病毒感染实验室小鼠 MHV-68（与KSHV密切相关）提供了高度易于处理的模型 人类γ-疱疹病毒的持久性，但该系统中的病毒潜伏期已有 在分子水平上没有充分表征以启用 出于这些目的对该模型的开发。因此，直接目标 拟议的研究是定义MHV-68的分子特征 潜伏期。为了获得我们的目标，我们提出了三个具体目标。在目标1下 我们将定义MHV-68潜伏基因表达的程序 造血细胞内潜伏期的建立和维护阶段 潜在病毒的储层（激活和静止的B细胞，巨噬细胞和 树突状细胞）。具体而言，我们将通过RT-PCR筛选这些细胞，并 MHV-68潜伏基因M2，M3的表达的Southern印迹杂交 M1 1、72、73和74。此外，我们建议识别潜在的新颖 这些细胞中表达的MHV-68潜伏基因从 病毒Genornic DNA序列的分析。在目标2下，我们将解决 三个延迟相关基因的功能：M2、73和74，其中很少或 没有信息可用。在AIM 3下，我们将定义MHV-68的影响 细胞基因表达在细胞水平上的潜伏期。一起， 这些具体目标将填补当前仍然存在于我们的显着空白 了解MHV-68的潜在生命周期，并提供更好的见解 进入γ-疱疹病毒作为一组维护的机制 潜伏期，因此有助于其致病潜力。