GLYCOPROTEIN B AND CMV INFECTION

人巨细胞病毒(Hcmv)是一种条件致病菌，可引起 免疫缺陷个体的多种疾病表现。本征 与巨细胞病毒感染相关的广泛的临床症状是 病毒感染不同类型和不同类型细胞的能力 发育谱系包括上皮细胞、内皮细胞、神经元和 血细胞。巨细胞病毒囊膜糖蛋白是人类免疫缺陷病毒 细胞嗜性与病毒诱导的融合反应的介体 对病毒渗透和细胞间传播负有责任 感染。拟议研究的目标是评估功能 关键的包膜糖蛋白gB在病毒进入宿主中的作用 细胞。为此，将采用两种相辅相成的方法。在……里面 基于基因的研究，我们将质疑这种缺失的后果 通过产生一种病毒株来减少这种丰富的关键包膜成分 缺乏GB蛋白质。目前正在生产一种gb阴性病毒株。 自从一种新型的永生化成纤维细胞系支持 人巨细胞病毒感染和空斑形成是在我们实验室进行的。我们 将对野生型的入门属性进行详细分析 一种含有完整包膜蛋白的病毒，而不是一GB 含有除gB外的所有包膜蛋白的缺陷病毒株。 具体活动将由于缺席或不参加而归于GB 具有GB负值的进入级联中特定事件的抑制 病毒。为了补充遗传学研究，生化特征 将进行重组衍生的、纯化的GB。精制产品的回收 保持天然构象和三级结构的蛋白质 为功能研究提供了一个极其有用的工具。使用这个 蛋白质，多个标准将被测试，以进一步探索其存在 以及GB的细胞受体的性质。在赠款的最后阶段， 我们将通过生产和生产GB来启动结构/功能分析 一组连接子插入突变体的特征。人类巨细胞病毒进入中国 宿主细胞是一个动态的过程，可能需要合作 多种病毒和细胞成分的参与。认识到 提出的研究目标将提供新的信息，不仅在 Gb在巨细胞病毒进入宿主细胞中的功能作用，但我们也将 获得更好的视角和更广泛的理解， 一种重要的人类病原体感染的关键阶段。

Human cytomegalovirus (HCMV) is an opportunistic pathogen that causes diverse disease manifestations in immune deficient individuals. Intrinsic to the broad range of clinical syndromes associated with HCMV infection is the ability of the virus to infect cells of distinct and divergent developmental lineages including, epithelial, endothelial, neuronal, and blood cells. HCMV envelope glycoproteins are the initial determinants of cellular tropism and the mediators of virally-induced fusion reactions that are responsible for virus penetration and cell-to-cell spread of infection. The goal of the proposed studies is to assess the functional role of a crucial envelope glycoprotein, gB, in virus entry into host cells. Towards this end, two complementary approaches will be applied. In genetically-based studies, we will question the consequences of the loss of this abundant, key envelope component by producing a strain of virus lacking a gB protein. Production of a gB-negative virus strain is now achievable since a novel, immortalized fibroblast cell line that supports HCMV infection and plaque formation was engineered in our laboratory. We will conduct a detailed analysis of the entry properties of the wild type virus that contains a full complement of envelope proteins versus a gB- deficient virus strain that contains all envelope proteins but gB. Specific activities will be attributed to gB by virtue of the absence or inhibition of a particular event in the entry cascade with the gB-negative virus. To complement the genetic studies, biochemical characterization of a recombinant-derived, purified gB will be performed. Recovery of purified protein that retains native conformation and tertiary structure has yielded an extremely useful tool for functional studies. Using this protein, multiple criteria will be tested to further explore the existence and nature of a cellular receptor for gB. In the final stage of the grant, we will initiate structure/function analysis of gB by producing and characterizing a panel of linker-insertion mutants. Entry of HCMV into host cells is a dynamic process likely requiring the cooperative participation of multiple viral and cellular components. The realization of the proposed research goals will provide novel information not only on the functional role of gB in HCMV entry into host cells but we will also gain a greater perspective and broader comprehension of this composite and critical stage in infection of a significant human pathogen.