VARICELLA-ZOSTER VIRUS MEMBRANE ANTIGENS

水痘-带状疱疹病毒是S嗜神经性人类疱疹病毒 这会导致两种不同的临床疾病：水痘(水痘) 带状疱疹(带状疱疹)。传播的、危及生命的威胁 VZV感染导致白血病儿童发生 目前正在接受临床治疗的VZV减毒株 评估。VZV以强烈的细胞相关方式复制， 从培养物中产生极低滴度的传染性病毒 细胞。尽管它作为人类病原体很重要，但我们只有 对生物、生物化学和 VZV的结构，但VZV有许多方面 很可能与病毒粒子-包膜相关的生物学。长的- 这项提案的学期目标是分析生物合成和 组装VZV信封并确定角色(S) 感染和免疫中的病毒包膜成分 对感染的反应。本提案中描述的实验 被设计成：(1)测试异常的蛋白水解性切割是否 GpII的表达与病毒感染性低有关；(2)定位 与单纯疱疹病毒(HSV)交叉反应的gpII表位 GB；(3)比较GPI的合成和加工动力学。 感染和未感染的细胞；(4)确定何时靶向 病毒糖蛋白对病毒粒子被膜核部位的影响 组装开始；(5)定位GPI的磷酸化位点； (6)分析预测的VZV蛋白激酶在VZV中的作用。 GPI的磷酸化；(7)分析了GPI的合成、加工和 次要糖蛋白gpIV的亚细胞定位 预测的糖蛋白GPV。所采用的技术将 包括：抗MAR单抗的制备 VZV突变体及其异型病毒粒子的制备 单纯疱疹病毒gB，免疫沉淀，SDS-PAGE，脉冲标记， 亚细胞分离、DNA克隆和测序、多肽 测绘，多肽特异性抗体的制备，Western 免疫印迹技术及表达非N蛋白的单纯疱疹病毒株的制备 单纯疱疹病毒糖蛋白。这些研究将产生试剂和 需要了解的信息 病毒糖蛋白及其翻译后修饰 病毒粒子组装和病毒感染，用于未来分析 对感染的免疫反应，以及可能发生的 不含DNA的亚单位疫苗。

Varicella-zoster virus (VZV) is s neurotropic human herpesvirus that causes two clinically distinct diseases: varicella (chickenpox) and zoster (shingles). The threat of disseminated, life-threatening VZV infections in leukemic children has led to the development of attenuated strains of VZV which are currently undergoing clinical evaluation. VZV replicates in a strongly cell-associated manner, and yields extremely low titers of infectious virus from cultured cells. Despite its importance as a human pathogen, we have only an incomplete understanding of the biology, biochemistry, and structure of VZV, but there are a number of aspects of VZV biology which are likely to be virion-envelope related. The long- term goals of this proposal are to analyze the biosynthesis and assembly of the VZV envelope and to determine the role(s) of the viral envelope components in infection and in the immune response to infection. The experiments described in this proposal are designed to: (1) test whether the unusual proteolytic cleavage of gpII is related to the low viral infectivity; (2) localize the epitopes on gpII that cross-react with herpes simplex virus (HSV) gB; (3) compare the kinetics of gpI synthesis and processing in infected and uninfected cells; (4) determine when targeting of the viral glycoproteins to the nuclear sites of virion envelope assembly begins; (5) locate the sites of the phosphorylation of gpI; (6) analyze the role of a predicted VZV protein kinase in the phosphorylation of gpI; (7) analyze the synthesis, processing and subcellular localization of the minor glycoprotein gpIV and of the predicted glycoprotein gpV. The techniques employed will include: preparation of monoclonal antibody resistant (MAR) mutants of VZV, preparation of heterotype VZV virions containing HSV gB, immunoprecipitation, SDS-PAGE, pulse-labelling, subcellular fractionation, DNA cloning and sequencing, peptide mapping, preparation of peptide-specific antibodies, Western immunoblotting, and preparation of HSV strains expressing non- HSV glycoproteins. These studies will yield reagents and information that are necessary for understanding the roles of the viral glycoproteins and their post-translational modifications in virion assembly and viral infection, for future analysis of the immune response to infection, and for possible development of a DNA-free subunit vaccine.