Murine Model of Gammaherpesvirus Latency

描述（由申请人提供）：拟议的长期目标 研究的目的是确定使伽马疱疹病毒能够 在具有免疫能力的宿主体内无限期地存在。这些机制 是与这些相关的发病机制和致癌潜力的基础 病毒，特别是在获得性或遗传性的个体中 免疫缺陷。不幸的是，由于极端的主机限制 人类 γ-疱疹病毒 EB 病毒 (EBV) 和卡波西病毒 肉瘤相关疱疹病毒（KSHV），目前还不可能直接 解决有关 EBV 和 KSHV 持续存在的机制 主机设置。实验小鼠感染鼠伽马疱疹病毒 MHV-68（与 KSHV 密切相关）提供了一种高度易于处理且具有潜力的模型 人类伽玛疱疹病毒的持久性，但该系统中的病毒潜伏期 尚未在分子水平上得到充分表征，以使得 利用该模型来达到这些目的。因此，近期目标 拟议研究的目的是确定 MHV-68 的分子特征 延迟。为了实现我们的目标，我们提出了三个具体目标。目标 1 下 我们将定义 MHV-68 潜伏期基因表达的程序 造血细胞内潜伏期的建立和维持阶段 潜伏病毒的储存库（激活的和静息的 B 细胞、巨噬细胞和 树突状细胞）。具体来说，我们将通过 RT-PCR 筛选这些细胞， Southern 印迹杂交用于表达 MHV-68 潜伏基因 M2、M3、 M1 1、72、73 和 74。此外，我们建议识别潜在的新颖性 MHV-68 潜伏基因在这些细胞中表达，但从 病毒基因组DNA序列分析。在目标 2 下，我们将解决 三个潜伏期相关基因的功能：M2、73 和 74，其中很少或 无可用信息。在目标 3 下，我们将定义 MHV-68 的影响 在细胞水平上体内细胞基因表达的潜伏期。在一起， 这些具体目标将填补我们目前仍然存在的重大差距 了解 MHV-68 的潜在生命周期，并提供更好的见解 研究伽马疱疹病毒作为一个群体利用维持的机制 潜伏期，从而有助于其致病潜力。

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.