VARICELLA-ZOSTER VIRUS MEMBRANE ANTIGENS

水痘-带状疱疹病毒(VZV)是一种人类疱疹病毒，是病原体 两种临床上不同的疾病：水痘(水痘)和带状疱疹 (带状疱疹)。尽管在正常个体中很少有生命危险，但主要的 (水痘)和再激活(带状疱疹)感染是 正在接受免疫抑制的癌症和器官移植患者 心理治疗。在许多情况下，VZV感染需要改变 化疗方案，以最大限度地减少播散性， 危及生命的感染。因为VZV的严重后果 这些免疫功能低下的患者受到感染，临床试验正在进行中 在美国和日本，为了测试活体免疫的效果， VZV减毒株。尽管它作为人类病原体很重要，但我们 对生物、生物化学和 VZV的结构，以及对感染的免疫反应。是这样的 信息是最终全面了解 VZV致病机制及康复机制的探讨 以及重新激活。本提案中描述的实验包括 旨在获得有关VZV包膜糖蛋白的基本信息， 免疫反应的可能目标。完整的鼠标面板 抗VZV糖蛋白的单抗，可在本 实验室，将被用于：(1)分析翻译后 VZV糖蛋白的修饰(O-连接糖基化， 棕榈化、硫酸盐化、磷酸化和蛋白分解)；(2)检查 糖蛋白的生物合成和加工；(3)纯化各糖蛋白 经典性和免疫亲和力测定的未变性糖蛋白 程序；以及(4)鉴定HSV和VZV糖蛋白，它们是 血清学交叉反应。纯化的糖蛋白将用于 分析中和抗体的VZV糖蛋白特异性 存在于VZV免疫球蛋白(VZIG)中。VZIG被动免疫是 临床上用于保护免疫功能低下的儿童免受严重、 危及生命的水痘。所采用的技术将包括： 免疫沉淀，十二烷基硫酸钠-PAGE，脉冲追逐标记，二维凝胶，部分 蛋白水解图、免疫印迹、免疫亲和层析 以及纸质和高效液相色谱(HPLC)。这些研究 将产生未来分析所需的试剂和信息 对VZV感染的免疫应答，为更好地发展 VZIG生产的筛选方法，将糖蛋白定位到 病毒基因组，以及可能开发的无DNA亚单位疫苗。

Varicella-zoster virus (VZV), a human herpesvirus, is the causative agent for two clinically distinct diseases: varicella (chickenpox) and shingles (zoster). Although rarely life-threatening in normal individuals, primary (varicella) and reactivated (zoster) infections are a major concern for cancer and organ transplant patients who are undergoing immunosuppressive therapy. In many cases, VZV infections necessitate changes in the chemotherapeutic regimen to minimize the risk of a disseminated, life-threatening infection. Because of the serious consequences of VZV infection in these immunocompromised patients, clinical trials are underway in the USA and Japan to test the efficacy of immunization with live, attenuated strains of VZV. Despite its importance as a human pathogen, we have only an incomplete understanding of the biology, biochemistry and structure of VZV, and of the immune response to infection. Such information is a prerequisite for the eventual full understanding of the pathogenesis of the diseases caused by VZV and the mechanisms of recovery and of reactivation. The experiments described in this proposal are designed to obtain basic information about the VZV envelope glycoproteins, the probable targets of the immune response. The complete panel of mouse monoclonal antibodies to the VZV glycoproteins, which is available in this laboratory, will be used to: (1) analyze the post-translational modifications of the VZV glycoproteins (O-linked glycosylation, palmitylation, sulfation, phosphorylation and proteolysis); (2) examine the biosynthesis and processing of the glycoproteins; (3) to purify each of the glycoproteins in undenatured form by classical and immunoaffinity procedures; and (4) identify the HSV and VZV glycoproteins which are serologically crossreactive. The purified glycoproteins will be used to analyze the VZV glycoprotein specificities of the neutralizing antibodies present in VZV-immune globulin (VZIG). Passive immunization with VZIG is used clinically to protect immunocompromised children from severe, life-threatening varicella. The techniques employed will include: immuno-precipitation, SDS-PAGE, pulse-chase labeling, 2-D gels, partial proteolytic mapping, Western immunoblotting, immunoaffinity chromatography and paper and high performance liquid chromatography (HPLC). These studies will yield reagents and information that are necessary for future analysis of the immune responses to VZV infection, for development of better screening methods for VZIG production, for mapping the glycoproteins to the viral genome, and for possible development of a DNA-free subunit vaccine.