STUDIES OF HERPES SIMPLEX VIRUS GLYCOPROTEINS

单纯疱疹病毒(HSV)是一种包膜DNA病毒，可导致人类 疾病，包括冻疮、眼睛和生殖器感染以及 脑炎。此外，生殖器疱疹可能在 艾滋病的获得者。病毒粒子的包膜含有七种糖蛋白， 它们在病毒感染中扮演着重要的角色，作为免疫的目标 宿主的反应，以及发病机制。这样做的长期目标是 格兰特是理解两种基本单纯疱疹病毒的三维结构 糖蛋白、Gd和Gh。目的1提出抗原性和抗原性研究 参与病毒穿透的蛋白质Gd的结构特性，以及 一种人类亚单位疫苗候选。我们结合了生化， 免疫学和重组DNA技术解决GD问题 结构，包括更精确地理解抗原表位， 二硫键的位置，N-键的作用 低聚糖，以及GD的低聚态。为了解释 积累的数据，必须知道GD的实际三维结构。 因此，目标2中的实验旨在分离出可结晶的 用于X射线结晶学三维结构测定的Gd形式。是这样的 研究将澄清智能交通系统中涉及的GD的结构元素 功能。该策略将通过以下步骤获得可结晶碎片 GDAIM 3蛋白水解物的抗原性和结构研究 一种参与病毒进入和细胞间的蛋白质Gh的性质 散开。将制备免疫试剂来对抗纯化的 蛋白质，并用于构建详细的抗原图。突变将会 被改造成克隆的Gh-1基因，突变蛋白将被 在转基因细胞中表达。突变体将被用于表位 地图绘制，并解决其他问题，如处理、运输和 功能。此外，我们将使用克隆的Gh-1基因来寻找Gh-2 同源同源。目标4提出了一项关于HSV糖蛋白潜在的研究 作为亚单位疫苗制剂并回答以下问题 问题。1)基因突变对GD的疗效有何影响 疫苗？2)其他单纯疱疹病毒糖蛋白能保护动物吗？3)其他单纯疱疹病毒 糖蛋白增强GD？的保护能力。

Herpes simplex viruses (HSVs) are enveloped DNA viruses that cause human disease, including cold sores, eye and genital infections and encephalitis. In addition, genital herpes may play a role in the acquisition of AIDS. The virion envelope contains seven glycoproteins, which play important roles in virus infection, as targets of the immune response of the host, and pathogenesis. The long term objective of this grant is to understand the 3-dimensional structure of two essential HSV glycoproteins, gD and gH. Aim 1 proposes studies of the antigenic and structural properties of gD, a protein involved in virus penetration, and a human subunit vaccine candidate. We have combined biochemical, immunologic and recombinant DNA techniques to address questions about gD structure, including a more precise understanding of antigenic epitopes, the position of disulfide bonds, the function of N-linked oligosaccharides, and the oligomeric state of gD. In order to interpret the accumulated data, the actual 3D structure of gD must be known. Therefore, experiments in Aim 2 are designed to isolate a crystallizable form of gD for 3-d structure determination by X-ray crystallography. Such studies would clarify the structural elements of gD involved in its functions. The strategy will be to obtain a crystallizable fragment by proteolysis of gD Aim 3 proposes studies of the antigenic and structural properties of gH, a protein involved in viral entry and cell-to -cell spread. Immunologic reagents will be prepared against the purified protein, and used to construct a detailed antigenic map. Mutations will be engineered into the cloned gH-1 gene, and mutant proteins will be expressed in transfected cells. The mutants will be used for epitope mapping, and to address other questions such as processing, transport and function. In addition we will use the cloned gH-1 gene to look for a gH-2 homolog. Aim 4 proposes a study of the potential of HSV glycoproteins to function as subunit vaccine preparations and to answer the following question. 1) What effect do mutations have on the efficacy of gD as a vaccine? 2) Can other HSV glycoproteins protect animals? 3) Do other HSV glycoproteins enhance the protective capacity of gD?