CYTOMEGALOVIRUS GENE REGULATION IN IMMUNODEFICIENCY

人巨细胞病毒(CMV)是影响人类健康的主要条件致病因子。 艾滋病患者。这种病毒被认为是几种可能的病毒之一 导致这种疾病进展的因素。病毒致病机制和 复制、持久性和延迟的决定因素是 巨细胞病毒在艾滋病患者体内的重新激活和传播，但机制 由于缺乏适当的模型系统，定义仍然不明确 这种物种特有的病毒。骨髓来源的造血细胞有 被认为是潜伏病毒的重要天然宿主。这 应用程序将识别能够容纳 病毒基因组，专注于粒细胞-巨噬细胞(GM)谱系，以及 将调查病毒基因表达的程度和病毒的形式 病毒基因组吸收了这些细胞。首先，我们将调查 GM系细胞在培养中作为病毒模型的静止性感染 研究已检测到的IE1基因在 这些细胞，鉴定其他病毒转录本，并分析 病毒基因组。这些信息将被用来指导调查 实验感染SCID-HU(骨/骨髓)小鼠和自然感染 来自血清阳性个体的受感染的骨髓细胞。第二，我们将 探讨病毒复活的信号(S)和机制(S) 被感染的GM前体细胞与PERMERED进行长期共培养 人成纤维细胞。刺激分化和增长的因素 将对转基因前体进行评估，以此作为提高以下方面效率的手段 重新激活。用于高效重新激活的重新激活参数 在培养的GM细胞模型和SCID-Hu小鼠中的病毒将应用于 自然感染者的外周血细胞或骨髓细胞。 长期目标是在生物测试中建立一个过程，该过程 从未屈服于实验性的操纵；病毒只显示出 当个人受到严重免疫抑制时重新激活，或当 潜伏感染的血液或组织被输注或移植到 另一个人。第三，我们将研究病毒的遗传基础 对于使用GM细胞培养的潜伏期(建立和重新激活) SCID-HU小鼠模型。个体调控基因(IE1、IE2)和新基因 已确定的潜伏感染相关基因将从病毒中删除 基因组和由此产生的突变病毒将被评估其 建立潜伏期和重新激活能力的生物学表型。 综上所述，拟议中的研究将定义生物光谱 在骨髓中的感染并将确定病毒的贡献 控制延迟和重新激活的功能。此信息将 提供有关病毒延迟的信息，这些信息应直接影响 控制免疫抑制后再激活的方法和 移植。

Human cytomegalovirus (CMV) is a major opportunistic pathogen affecting AIDS patients. This virus has been suggested to be one of several possible contributors to progression in this disease. Viral pathogenesis and determinants of replication, persistence and latency are key to reactivation and dissemination of CMV in AIDS patients but mechanisms remain poorly defined because of a lack of appropriate model systems for this species specific virus. Bone marrow derived hematopoietic cells have been thought to be an important natural reservoir of latent virus. This application will identify bone marrow cell types capable of harboring the viral genome, focussing on the granulocyte-macrophage (GM) lineage, and will investigate the extent of viral gene expression and the form that the viral genome takes in these cells. First, we will investigate the quiescent infection of the GM lineage cells in culture as a model of viral latency, studying the expression of the ie1 gene that has been detected in these cells, identifying other viral transcripts and analyzing the form of the viral genome. This information will be used to guide investigations in experimentally infected SCID-hu (bone/bone marrow) mice and naturally infected bone marrow cells from seropositive individuals. Second, we will investigate the signal(s) and mechanism(s) of viral reactivation when infected GM precursors are subjected to long-term coculture with permissive human fibroblast cells. Factors that stimulate differentiation and growth of GM precursors will be evaluated as a means to increase efficiency of reactivation. Parameters for reactivation that efficiently reactivate virus in the cultured GM cell model and in SCID-hu mice will be applied to peripheral blood or bone marrow cells of naturally infected individuals. The long term aim is to establish in biological assay for a process that has never yielded to experimental manipulation; virus has only been shown to reactivate when an individual is severely immunosuppressed, or when latently infected blood or tissues are transfused or transplanted into another human being. Third, we will investigate the viral genetic basis for latency (establishment and reactivation) using the GM cell culture and SCID-hu mouse models. Individual regulatory genes (ie1, ie2) and newly identified latent infection associated genes will be deleted from the viral genome and the resultant mutant viruses will be evaluated for their biological phenotype in establishment of latency and ability to reactivate. Taken together, the proposed research will define the biological spectrum of infection in bone marrow and will establish the contribution of viral functions that control latency and reactivation. This information will provide information on viral latency that should directly impact on the approaches to control reactivation following immunosuppression and transplantation.