VIRAL DETERMINANTS OF HSV DISEASE IN MICE

小鼠 HSV 疾病的病毒决定因素

• 批准号: 6099516
• 负责人: PATRICIA G SPEAR
• 金额: --
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至 1999-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6099516
• 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; 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.

该项目的远程目标是定义细胞和病毒 进入单纯疱疹病毒类型1和2的要求（HSV-1和 HSV-2）进入生殖道的细胞，以使感染传播 进入其他组织的门户，包括神经系统。这 直接目的是识别介导的HSV-1和HSV-2糖蛋白 病毒进入阴道上皮细胞，是 病毒从上皮到细胞的细胞间扩散所必需的 外周和中枢神经系统的。有限的信息 可用于HSV-1的感染培养细胞的需求 不能完全推广到HSV-2或分化的感染 完整宿主的细胞。为个体删除的突变病毒菌株 糖蛋白基因或表达糖蛋白的改变形式将 源自父母菌株HSV-1（F）和HSV-2（G）。父母 突变菌株将用于通过腔内接种小鼠 灌输。生殖道和神经系统的细胞 然后将被识别为被感染和病理变化， 包括中枢神经系统病变和脱髓鞘，与 感染。父母和突变的病毒菌株将表达β- 半乳糖苷酶在强IE巨细胞病毒启动子的控制下 来自单个基因间区域，因此受感染的细胞很容易成为 通过裂解产生的彩色产物在小鼠组织中鉴定 通过β-半乳糖苷酶的X-GAL。要删除的糖蛋白包括，包括 根据优先顺序，GB，GC，GD，GE，GI，GG，GH，GH和GL（一些 可能从另一个项目获得所需的删除突变体）。每个 这些糖蛋白与病毒结合或进入 各种类型的培养细胞，或介导细胞间扩散 感染。具有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补充细胞上传播 产物，这是在培养细胞中复制所必需的。两个都 可以对病毒的补充和未结合形式进行测试是否具有能力 感染小鼠。我们期望确定哪种病毒糖蛋白是 将病毒进入上皮细胞的顶部表面所需 那条阴道腔，上皮区域是 优先感染了哪种病毒糖蛋白需要哪种病毒 感染从入门细胞传播到其他细胞的其他细胞 上皮以及背根神经节和其他细胞中的神经元 在脊髓和大脑中。 这些结果，结果与结果 在豚鼠和培养细胞中的其他计划研究中获得 从人类的女性生殖道，应允许确定 女性生殖道感染的特定要求； 探索可以控制自然历史差异的因素 HSV-1和HSV-2引起的疾病和病理；和识别 对于传染性非 - 致病疫苗菌株。