STUDIES OF HERPES SIMPLEX VIRUS GLYCOPROTEINS

单纯疱疹病毒（HSV）是一种有包膜的DNA病毒， 疾病，包括唇疱疹，眼睛和生殖器感染， 脑炎生殖器疱疹的症状有哪些？ 获得艾滋病。病毒体包膜含有七种糖蛋白， 在病毒感染中起重要作用，作为免疫的靶点， 宿主的反应和发病机制。这一长期目标 格兰特是了解两个基本的HSV的三维结构 糖蛋白，gD和gH。目的1提出了抗原性和 gD（一种参与病毒穿透的蛋白质）的结构特性，以及 人亚单位疫苗候选物。我们结合了生物化学， 免疫学和重组DNA技术解决gD问题 结构，包括对抗原表位的更精确理解， 二硫键的位置，N-连接的功能 低聚糖和gD的低聚状态。为了诠释 积累的数据，gD的实际3D结构必须是已知的。 因此，目的2中的实验被设计为分离可结晶的 用于通过X射线晶体学确定三维结构的gD形式。等 研究将阐明参与其作用的gD的结构要素， 功能协调发展的该策略将是通过以下方式获得可结晶片段： 目的3提出了gD蛋白水解的抗原性和结构的研究， gH是一种参与病毒进入和细胞间相互作用的蛋白质， 传播.免疫试剂将针对纯化的 蛋白质，并用于构建详细的抗原图谱。突变将 将被工程化到克隆的gH-1基因中，突变蛋白将被 在转染的细胞中表达。突变体将用于表位分析。 测绘，并解决其他问题，如加工，运输和 功能此外，我们将利用克隆的gH-1基因寻找一个gH-2 同源物目的4：研究HSV糖蛋白对 作为亚单位疫苗制剂的功能，并回答以下问题 问题1)突变对gD作为一种抗肿瘤药物的疗效有什么影响？ 疫苗？2)其他HSV糖蛋白能保护动物吗？3)其他HSV 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?