CYTOMEGALOVIRUS GENE REGULATION IN IMMUNODEFICIENCY

人巨细胞病毒（CMV）是一种主要的机会致病菌， 艾滋病患者。 这种病毒被认为是 导致这种疾病的进展。 病毒发病机制和 复制、持久性和延迟的决定因素是 艾滋病患者中CMV的再激活和传播，但机制 由于缺乏适当的模型系统， 这种特定物种的病毒。 骨髓来源的造血细胞 被认为是潜伏病毒的重要天然储存库。 这 应用程序将确定骨髓细胞类型能够窝藏 病毒基因组，集中于粒细胞-巨噬细胞（GM）谱系，以及 将研究病毒基因表达的程度和病毒基因表达的形式。 病毒基因组进入这些细胞。 首先，我们将调查 作为病毒感染模型的培养物中GM谱系细胞的静止感染 潜伏期，研究已检测到的ie 1基因的表达， 这些细胞，识别其他病毒转录本，并分析 病毒基因组 这些信息将用于指导调查， 实验感染的SCID-hu（骨/骨髓）小鼠和自然感染的SCID-hu（骨髓）小鼠 来自血清反应阳性个体的感染骨髓细胞。 二是 研究病毒再激活的信号和机制， 受感染的转基因前体进行长期共培养， 人成纤维细胞 刺激分化和生长的因素 将评估转基因前体的使用情况，作为提高 重新激活 有效重新激活的重新激活参数 在培养的GM细胞模型和SCID-hu小鼠中的病毒将被应用于 自然感染个体的外周血或骨髓细胞。 长期目标是在生物测定中建立一种方法， 从未屈服于实验操作;病毒只被证明是 当个体严重免疫抑制时，或当 潜伏感染的血液或组织被输入或移植到 另一个人 第三，我们将调查病毒的遗传基础 使用GM细胞培养物进行潜伏期（建立和再激活）， SCID-hu小鼠模型。 个别调控基因（ie 1，ie 2）和新的 鉴定的潜伏感染相关基因将从病毒中删除， 将评估基因组和所得突变病毒的 在建立潜伏期和重新激活能力方面的生物表型。 综合考虑，拟议中的研究将定义生物光谱 并将确定病毒的贡献 控制延迟和重新激活的功能。 这些信息将 提供有关病毒潜伏期的信息，这些信息应直接影响 控制免疫抑制后再活化的方法， 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.