VARICELLA-ZOSTER VIRUS MEMBRANE ANTIGENS

水痘-带状疱疹病毒（VZV），一种人类疱疹病毒，是致病因子 两种临床上不同的疾病：水痘和带状疱疹 （带状疱疹）。 虽然在正常人中很少危及生命，但原发性 （水痘）和再激活（带状疱疹）感染是一个主要问题， 正在接受免疫抑制治疗的癌症和器官移植患者 疗法 在许多情况下，VZV感染需要改变 化疗方案，以最大限度地减少扩散的风险， 危及生命的感染 由于VZV的严重后果 在这些免疫功能低下的患者中，临床试验正在进行中 在美国和日本， VZV减毒株。 尽管它作为人类病原体的重要性，我们 我对生物学、生物化学和 VZV的结构和对感染的免疫反应。 等 信息是最终充分了解 VZV引起的疾病的发病机制和康复机制 和重新激活。 本提案中描述的实验是 旨在获得关于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 VZV糖蛋白的单克隆抗体，这是在这个 实验室，将用于：（1）分析翻译后 VZV糖蛋白的修饰（O-连接的糖基化， 棕榈酰化，硫酸化，磷酸化和蛋白水解）;（2）检查 （3）纯化糖蛋白的每一种， 通过经典和免疫亲和法测定未变性形式的糖蛋白 程序;和（4）鉴定HSV和VZV糖蛋白 血清交叉反应 纯化的糖蛋白将用于 分析中和抗体的VZV糖蛋白特异性 VZV免疫球蛋白（VZIG）。 VZIG被动免疫是 临床上用于保护免疫功能低下的儿童， 危及生命的水痘 所采用的技术将包括： 免疫沉淀，SDS-PAGE，脉冲追踪标记，2-D凝胶，部分 蛋白酶图谱，免疫印迹，免疫亲和层析 以及纸和高效液相色谱法（HPLC）。 这些研究 将产生未来分析所需的试剂和信息 的免疫反应VZV感染，为发展更好的 用于VZIG生产的筛选方法，用于将糖蛋白定位到VZIG的糖蛋白上， 病毒基因组，并可能开发无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.