ANTIGENIC DETERMINANTS OF VARICELLA VIRUS

水痘-带状疱疹病毒（VZV）是两种常见的病原体， 儿童疾病水痘，并在重新激活后， 带状疱疹（herpes zoster） 虽然只有适度的痛苦， 健康的人，这两种疾病是发病的重要原因， 儿童和成人癌症或其他慢性 免疫系统疾病，包括艾滋病。 研究的目的是 为了定义和表征这种病毒的主要抗原决定簇， 疱疹病毒 由VZV编码的糖蛋白将被给予最高的 优先考虑，因为它们在诱导保护性 免疫反应;相同的糖蛋白介导重要的相互作用 在病毒和细胞之间，也作为细胞表面受体发挥作用。 VZV 在人类疱疹病毒中是独一无二的，具有最少的糖蛋白; 本建议的重点将限于这两种糖蛋白 在VZV基因组的Us片段内编码：VZV gpI和gpIV。 的 具体目的包括：（1）分析细胞表面 VZV诱导Fc受体的表达和糖蛋白基因的制备 模拟其生物学活性的构建体;（2）研究细胞 催化VZV gpI和gpIV磷酸化的蛋白激酶;（3） 克隆和表达两种推定的VZV蛋白激酶， 它们是否磷酸化两种糖蛋白;以及（4）定义 VZV Fc受体和该组成糖蛋白的功能。 gpI和gpIV的表位参与了抗体依赖性细胞免疫应答。 VZV感染细胞的毒性和抗体加补体溶解， 病毒Fc受体结合后可能被抑制的活性 通过非免疫IgG或双极抗体桥接。 方法包括亚克隆 用于转录/翻译和转染的VZV糖蛋白基因 实验;用细胞磷酸转移酶进行的蛋白激酶测定;位点- 丝氨酸/苏氨酸残基的定向诱变; 假定的病毒激酶;杂交瘤生产和流式细胞术，以及 功能测定以表征抗原位点。 从这些 研究应明确两种VZV的主要结构-功能关系 我们的糖蛋白，同时，提供至少部分 解释为什么VZV可以是一种有能力的α/疱疹病毒， 糖蛋白的数量。

Varicella-zoster virus (VZV) is the etiologic agent of both the common childhood illness chickenpox and, upon reactivation, the dermatomal exanthem shingles (herpes zoster). Although only moderately distressful in healthy people, both diseases are important causes of morbidity and mortality in children and adults with cancer or other chronic immunocompromising illnesses, including AIDS. The goal of the research is to define and characterize the major antigenic determinants of this herpesvirus. Glycoproteins encoded by VZV will be given the highest priority because they play a prominent role in the induction of protective immune responses; the same glycoproteins mediate important interactions between virus and cell, and also function as cell surface receptors. VZV is unique among the human herpesviruses in having the fewest glycoproteins; the focus of this proposal will be restricted to the two glycoproteins encoded within the Us segment of the VZV genome: VZV gpI and gpIV. The Specific Aims include the following: (1) Analyze the cell surface expression of a VZV induced Fc receptor and prepare glycoprotein gene constructs which mimic its biologic activity; (2) Investigate the cellular protein kinases which catalyze phosphorylation of VZV gpI and gpIV; (3) Clone and express the two putative VZV protein kinases and determine whether they phosphorylate the two glycoproteins; and (4) Define the functions of the VZV Fc receptor and this constituent glycoproteins. Epitopes of gpI and gpIV are involved in antibody dependent cellular toxicity and antibody plus complement lysis of VZV-infected cells, activities which may be inhibited after engagement of the viral Fc receptor by nonimmune IgG or bipolar antibody bridging. Methods include subcloning of VZV glycoprotein genes for transcription/translation and transfection experiments; protein kinase assays with cellular phosphotransferases; site- directed mutagenesis of serine/threonine residues; analyses of cloned putative viral kinases; hybridoma production and flow cytometry, as well as functional assays to characterize antigenic sites. Results from these studies should define major structure-function relationships of the two VZV Us glycoproteins, and at the same time, provide at least a partial explanation why VZV can be a competent alpha/herpesvirus with a limited number of glycoproteins.