VIRAL DETERMINANTS OF HSV DISEASE IN MICE

小鼠 HSV 疾病的病毒决定因素

• 批准号: 6235005
• 负责人: PATRICIA G SPEAR
• 金额: $ 13.96万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-07-01 至 1998-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6235005
• 关键词: cooperative study; dorsal root; female; ganglions; gene deletion mutation; genetic strain; glycoproteins; guinea pigs; herpes simplex virus 1; herpes simplex virus 2; laboratory mouse; molecular pathology; mutant; myelination; nervous system infection; tissue /cell culture; vagina; virulence; virus infection mechanism; virus protein

The long-range goals of this project are to define cell and viral requirements for entry of herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) into cells of the genital tract and for spread of infection from the portal of entry to other tissues, including the nervous system. The immediate aims are to identify HSV-1 and HSV-2 glycoproteins that mediate the entry of virus into cells of the vaginal epithelium and that are required for the cell-to-cell spread of virus from the epithelium to cells of the peripheral and central nervous system. The limited information available about requirements for infection of cultured cells by HSV-1 may not be fully generalizable to HSV-2 or to the infection of differentiated cells of the intact host. Mutant viral strains deleted for individual glycoprotein genes, or expressing altered forms of the glycoprotein, will be derived from the parental strains HSV-1 (F) and HSV-2(G). The parental and mutant strains will be used to inoculate mice by intravaginal instillation. The cells of the genital tract and nervous system that become infected will then be identified and pathological changes, including CNS lesions and demyelination, correlated with the-spread of infection. The parental and mutant viral strains will express beta- galactosidase under control of the strong IE cytomegalovirus promoter, from a single intergenic region, so that infected cells can readily be identified in mouse tissues by the colored product resulting from cleavage of X-gal by beta-galactosidase. The glycoproteins to be deleted include, in order of priority, gB, gC, gD, gE, gI, gG, gH, and gL (some of the desired deletion mutants may be available from another project). Each of these glycoproteins has been implicated in viral binding to or entry into cultured cells of various types, or in mediating cell-to-cell spread of infection. Viral mutants with deletions of the gB, gD, gH and gL genes must be propagated on complementing cells that provide the missing gene product, which is required for replication in cultured cells. Both complemented and noncomplemented forms of virus can be tested for ability to infect mice. We expect to determine which viral glycoproteins are required for entry of virus into the apical surfaces of epithelial cells that line the vaginal cavity, which regions of the epithelium are preferentially infected, which viral glycoproteins are required for the spread of infection from the portal-of-entry cells to other cells of the epithelium and also to neurons in dorsal root ganglia and to other cells in the spinal cord and brain. These results, considered with results obtained in other planned studies in guinea pigs and in cells cultured from the human female genital tract, should allow determination of specific requirements for infection of the female genital tract; exploration of factors that can govern differences in natural history of disease and pathology caused by HSV-1 and HSV-2; and identification of mutations that would be desirable for the engineering of infectious non- pathogenic vaccine strains.

这个项目的长期目标是定义细胞和病毒 单纯疱疹病毒1型和2型(HSV-1和HSV-2)入境要求 HSV-2)进入生殖道细胞，并从 进入其他组织的入口，包括神经系统。这个 目前的目标是识别HSV-1和HSV-2糖蛋白 病毒进入阴道上皮细胞，这是 病毒从上皮到细胞的细胞间传播所必需的 外周和中枢神经系统。有限的信息 关于HSV-1感染培养细胞的要求可能 不能完全概括到HSV-2或分化型感染 完整宿主的细胞。为个人删除突变病毒株 糖蛋白基因，或表达改变形式的糖蛋白，将 来源于亲本株HSV-1(F)和HSV-2(G)。父母 突变株将通过阴道接种小鼠 灌输。生殖道和神经系统的细胞 被感染的人将被识别并发生病理变化， 包括中枢神经系统损害和脱髓鞘，与肿瘤的扩散相关 感染。亲本和突变的病毒株将表达β- IE巨细胞病毒强启动子控制下的半乳糖苷酶， 来自单个基因间隔区，因此感染的细胞可以很容易地 通过卵裂产生的有色产物在小鼠组织中进行鉴定 β-半乳糖苷酶对X-半乳糖的影响。要删除的糖蛋白包括， 按照优先级顺序，GB、GC、Gd、Ge、Gi、Gg、Gh和gl(一些 可以从另一个项目获得所需的删除突变体)。每一个 这些糖蛋白被认为与病毒结合或进入 培养的各种类型的细胞，或在介导细胞间的传播 感染。Gb、Gd、Gh和gl基因缺失的病毒突变体 必须在提供缺失基因的互补细胞上繁殖 产品，这是在培养细胞中复制所需的。两者都有 可以测试补体和非补体形式的病毒的能力 用来感染老鼠。我们希望确定哪些病毒糖蛋白是 病毒进入上皮细胞根尖表面所必需的 阴道腔的线条，上皮的哪些区域 优先感染的，哪些病毒糖蛋白是需要的 感染从入口细胞向其他细胞的扩散 上皮细胞以及背根神经节中的神经元和其他细胞 在脊髓和大脑中。这些结果，与结果一起考虑 在其他计划中的豚鼠和细胞培养研究中获得 来自人类女性生殖道的，应该允许测定 女性生殖道感染的具体要求； 自然历史差异的制约因素探讨 HSV-1和HSV-2引起的疾病和病理；以及 突变，这将是理想的工程传染性非 致病疫苗毒株。