MOLECULAR GENETICS OF VARICELLA ZOSTER VIRUS

水痘带状疱疹病毒的分子遗传学

• 批准号: 2566856
• 负责人: J I COHEN
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2566856
• 关键词: beta galactosidase; disease /disorder model; gene mutation; genetic manipulation; glycoproteins; guinea pigs; herpes simplex virus 1; herpes simplex virus 2; latent virus infection; molecular genetics; tissue /cell culture; transfection; uveitis; varicella zoster virus; virus genetics; virus infection mechanism; virus protein; virus replication

Varicella-zoster virus (VZV) is the etiologic agent of chickenpox and herpes zoster. We have developed a system to mutate individual genes or to insert foreign DNAs in the VZV genome. These mutant viruses are being assayed for their ability to grow in cell culture or in animals models. Deletion or inactivation of some of these genes (ORF1, ORF8, ORF10, ORF13, ORF14, ORF47, ORF66) results in no discernable difference in the growth of the virus in cell culture, while inactivation of other genes (ORF9A, ORF19) or pairs of genes (ORF 47 and ORF66) reduces the ability of the virus to grow in cell culture. Analysis of VZV mutants has shown that VZV ORF47 and ORF66 have protein kinase activity and that ORF9A is important for syncytia formation of VZV-infected cells. Guinea pigs will be inoculated with selected VZV mutants to determine if the viruses have lost the ability to establish latency or reactivate from the central nervous system. Foreign genes have been inserted into the VZV genome. Intravitreal inoculation of guinea pigs with VZV expressing beta-galactosidase resulted in a chronic uveitis with a mononuclear cell infiltrate in the vitreous of nearly all of the animals that persisted for at least two months. Staining with X-gal demonstrated the presence of VZV in the ciliary body or iris in about 50% of the animals for at least two months after inoculation. Inoculation of guinea pigs with VZV expressing herpes simplex virus glycoprotein D, followed by challenging the animals with intravaginal herpes simplex virus, resulted in reduced severity of genital herpes.

水痘-带状疱疹病毒（VZV）是水痘的病原体， 带状疱疹。 我们已经开发出一种系统， 将外源DNA插入VZV基因组 这些变异病毒 测定它们在细胞培养物或动物模型中生长的能力。 这些基因中的一些（ORF 1，ORF 8，ORF 10， ORF 13、ORF 14、ORF 47、ORF 66）在细胞内的表达没有可辨别的差异。 病毒在细胞培养中的生长，而其他基因的失活 （ORF 9A，ORF 19）或成对的基因（ORF 47和ORF 66）降低了 在细胞培养中生长。 对VZV突变体的分析表明， VZV ORF 47和ORF 66具有蛋白激酶活性，ORF 9A 对VZV感染细胞的合胞体形成很重要。 豚鼠将 接种选定的VZV突变体，以确定病毒是否具有 失去了建立延迟或从中央重新激活的能力 神经系统 外源基因已被插入VZV基因组中。 玻璃体 用表达β-半乳糖苷酶的VZV接种豚鼠 导致慢性葡萄膜炎，单核细胞浸润， 几乎所有动物的玻璃体，持续至少两个 个月 用X-gal染色证明VZV在细胞中的存在。 睫状体或虹膜中的约50%至少两个月 接种后。用表达疱疹的VZV接种豚鼠 单纯病毒糖蛋白D，然后用 阴道内单纯疱疹病毒，导致严重程度降低 生殖器疱疹