ENTRY OF HERPES SIMPLEX VIRUS INTO CELLS

这里提出的研究的最终目标是定义 单纯疱疹病毒(HSV)侵袭人类的机制 从而启动病毒基因的表达。目前的努力是 旨在识别和表征细胞表面成分 影响HSV与细胞和条目的结合以及对定义 介导HSV结合和HSV结合的HSV糖蛋白功能域 进入。HSV与细胞的最初接触是通过结合到 细胞表面蛋白多糖(PG)的糖胺多聚糖链(GAG)。这个 病毒粒子糖蛋白GC主要负责这种结合 而其他病毒粒子糖蛋白(Gb、Gd、Gh和Gl)则需要 穿透力。大多数培养的细胞对HSV感染都很敏感。一 例外的是中国仓鼠卵巢(CHO)细胞系，它是 部分抵抗某些HSV株的进入，如HSV-1(KOS)，但 完全受其他人的影响。为了更好地定义细胞表面 影响HSV结合和进入的组件，我们打算(I) 确定可作为受体的特定PG(至少部分 通过其Gag链)用于HSV与细胞的结合；(Ii)探索 此外，一些初步证据表明，GAG可能会激活对 HSV，以及作为病毒结合的受体和(Iii) 鉴定可使野生型和突变型CHO细胞 容易被细胞感染的单纯疱疹病毒株感染 抵抗力强。为了更好地定义HSV糖蛋白的功能结构域， 介导HSV感染，我们打算(Iv)分离和鉴定HSV 表现出特异性改变的单纯疱疹病毒突变体的基因改变 细胞结合；(V)产生重组形式的HSV糖蛋白 介导病毒结合，以探索细胞表面 病毒识别的配体的表达和分布 野生型和变异型特异性糖蛋白及(V)测定 有能力感染CHO细胞的病毒的哪个或哪些基因必须 被转移到HSV-1(KOS)，以便使HSV-1(KOS)能够胜任 感染，从而识别表现细胞类型的可变HSV基因- 依赖于它们调节病毒进入的能力。 这些研究的结果应该更好地确定细胞表面的PG HSV结合的糖蛋白和病毒糖蛋白的功能结构域 调停捆绑。此外，还需要其他细胞表面组件 可能被确定为HSV进入和控制HSV进入的病毒基因 将确定特定于细胞的时尚。信息和探查 在本研究过程中开发的将允许更全面的 那么，现在可以对一些关键的决定因素进行调查 单纯疱疹病毒的发病机制。新型抗病毒药物和治疗性或预防性药物 更好地了解细胞的机制可能会产生策略 单纯疱疹病毒的入侵。

The ultimate objective of studies proposed here is to define the mechanism or mechanisms by which herpes simplex virus (HSV) invades a cell so as to initiate viral gene expression. Current efforts are directed toward identifying and characterizing cell surface components that influence HSV binding to cells and entry and also toward defining functional domains of the HSV glycoproteins that mediate HSV binding and entry. HSV makes its initial contact with cells by binding to glycosaminoglycan (GAG) chains of cell surface proteoglycans (PGs). The virion glycoprotein gC is principally responsible for this binding whereas other virion glycoproteins (gB, gD, gH and gL) are required for penetration. Most cultured cells are susceptible to HSV infection. One exception is the Chinese hamster ovary (CHO) cell line, which is partially resistant to entry of some HSV strains, such as HSV-1(KOS), but is fully susceptible to others. To better define the cell surface components that influence HSV binding and entry, we intend (i) to identify specific PGs that can serve as receptor (at least in part through their GAG chains) for HSV binding to cells; (ii) to explore further some preliminary evidence that GAGs may activate penetration of HSV, as well as serve as receptors for virus binding and (iii) to identify human genes that can render wild-type and mutant CHO cells susceptible to infection by strains of HSV to which the cells are resistant. To better define functional domains of HSV glycoproteins that mediate HSV infection, we intend to (iv) to isolate and characterize the genetic alterations of HSV mutants that exhibit altered specificity for cell binding; (v) to produce recombinant forms of the HSV glycoproteins that mediate virus binding, in order to explore the cell surface expression and distribution of ligands recognized by the viral glycoproteins with wild-type and altered specificity and (v) to determine which gene or genes of a virus that is competent to infect CHO cells must be transferred to HSV-1(KOS) in order to render HSV-1(KOS) competent for infection, thus identifying variable HSV genes that exhibit cell type- dependence in their ability to mediate viral entry. The results of these studies should better define the cell surface PGs to which HSV binds and the functional domains of viral glycoproteins that mediate the binding. In addition, other cell surface components required for HSV entry may be identified and viral genes that govern entry in a cell-specific fashion will be identified. The information and probes developed during the course of this study will permit a more thorough investigation, then is now possible, of some of the key determinants of HSV pathogenesis. Novel antiviral drugs and therapeutic or prophylactic strategies could result from better understanding the mechanism of cell invasion by HSV.