VARICELLA-ZOSTER VIRUS MEMBRANE ANTIGENS

水痘-带状疱疹病毒（VZV）是一种嗜神经的人类疱疹病毒 导致两种临床上不同的疾病：水痘（水痘） 和带状疱疹。 扩散的威胁，危及生命的威胁 白血病儿童的VZV感染导致了 目前正在进行临床试验的VZV减毒株 评价 VZV以强烈的细胞结合方式复制， 并从培养的病毒中产生极低滴度的感染性病毒， 细胞 尽管它作为人类病原体的重要性， 对生物学、生物化学和 VZV的结构，但VZV有许多方面 可能与病毒体包膜相关的生物学。 很长的- 该提案的长期目标是分析生物合成和 组装VZV信封，并确定 感染和免疫中的病毒包膜成分 对感染的反应。 本提案中所述的实验 目的是：（1）测试不寻常的蛋白水解切割是否 gpII的表达与病毒感染性低有关;（2）定位gpII的表达， gpII上与单纯疱疹病毒（HSV）交叉反应的表位 gB;（3）比较gpI合成和加工的动力学， 感染和未感染的细胞;（4）确定何时靶向 病毒糖蛋白到病毒体包膜的核位点 组装开始;（5）定位gpI磷酸化位点; (6)分析预测的VZV蛋白激酶在 gpI的磷酸化;（7）分析合成，加工和 次要糖蛋白gpIV的亚细胞定位和 预测的糖蛋白gpV。 所采用的技术将 包括：制备抗MAR单克隆抗体 VZV的突变体、含有VZV的异型病毒粒子的制备 HSV gB，免疫沉淀，SDS-PAGE，脉冲标记， 亚细胞分级，DNA克隆测序，肽 定位，肽特异性抗体的制备，Western 免疫印迹和制备表达非- HSV糖蛋白 这些研究将产生试剂， 了解所需的信息 病毒糖蛋白及其翻译后修饰 病毒粒子组装和病毒感染，为未来的分析 对感染的免疫反应，以及可能发生的 无DNA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.