PROTECTION OF GENITAL MUCOSA AND GANGLIA AGAINST HSV-2

保护生殖器粘膜和神经节免受 HSV-2 感染

• 批准号: 6349855
• 负责人: Gregg N. Milligan
• 金额: $ 7.44万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-01 至 2001-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6349855
• 关键词: Herpesviridae vaccine; attenuated microorganism; cancer prevention; disease /disorder prevention /control; genital herpes; herpes simplex virus 2; laboratory mouse; microorganism culture; microorganism immunology; mucosa; mucosal immunity; neoplasm /cancer vaccine; thymidine kinase; vaccine development; vagina

Greater than Herpes simplex virus type 2 (HSV-2) now infects the genital tracts of approximately one in five Americans. Strategies to prevent new HSV-2 infections are complicated by its ability to initiate a latent infection in the sensory ganglia, periodically reactivate, and cause recurrent lesions or asymptomatic virus shedding in genital tissues thus increasing its chances of infecting new hosts. Effective vaccines are needed to prevent the establishment of latency within the sensory ganglia. Unfortunately, little is known about the immune mechanisms which protect the sensory ganglia. In animal models, previous genital inoculation with HSV-2 elicits immunity which protects the sensory ganglia from reinfection, thus serving as a paradigm for an effective HSV-2 vaccine. The long term aims of this proposal are to use a murine model of genital HSV-2 inoculation to understand the types of immune mechanisms responsible for protection, how these mechanisms work at the molecular level, and how to elicit these responses to provide long term protection. The results of these studies will provide important information for the rational design of vaccines to protect against HSV-2. In the first aim, a recombinant HSV-2 strain expressing green fluorescent protein (HSV-2 gfp) will be used as a marker to determine if HSV-specific T lymphocytes prevent HSV-2 from reaching the sensory ganglia, thus preventing the establishment of latency. Quantification of HSV-2 gfp infected ganglionic neurons by UV microscopy and HSV-2 gfp genomes in the ganglia by quantitative PCR will be used to demonstrate the role of specific T cell subsets in preventing acute and latent HSV-2 infection of the ganglia. In the second aim, an antibody deficient strain of mice (muMT) will be used to determine the role of HSV-specific antibody in protection of the sensory ganglia. Purified IgG and IgA fractions of HSV-specific sera will be administered to HSV-immune muMT mice to determine the efficacy of specific antibody isotypes in completing the protection of HSV-immune muMT mice against the establishment of latent HSV-2 infection. In the third aim, the ability of immune responses elicited by inoculation of distal mucosal or systemic sites to protect the sensory ganglia will be tested. The ability of inoculation at these sites to elicit long term memory immune responses within the vaginal mucosa and associated genital lymphoid tissue which can be rapidly recalled for protection of the vaginal mucosa and sensory ganglia will be assessed.

现在，比单纯疱疹病毒2型（HSV-2）感染了大约五分之一的美国人的生殖道。防止新的HSV-2感染的策略由于其在感觉神经节中引发潜在感染，定期重新激活并引起复发性病变或生殖器组织中脱落的复发性病毒的能力而变得复杂，从而增加了其感染新宿主的机会。 需要有效的疫苗来防止在感觉神经节内建立潜伏期。 不幸的是，对保护感觉神经节的免疫机制知之甚少。 在动物模型中，先前使用HSV-2的生殖器接种引起了保护感觉神经节免于再感染的免疫力，因此可以作为有效HSV-2疫苗的范例。 该提案的长期目的是使用生殖器HSV-2接种的鼠模型来了解负责保护的免疫机制的类型，这些机制在分子水平上的工作方式以及如何引起这些反应以提供长期保护。 这些研究的结果将为疫苗的合理设计提供重要信息，以防止HSV-2。在第一个目标中，将使用表达绿色荧光蛋白（HSV-2 GFP）的重组HSV-2菌株用作标记，以确定HSV特异性T淋巴细胞是否阻止HSV-2阻止HSV-2达到感觉神经节，从而防止了潜伏期的建立。 通过定量PCR在神经节中对HSV-2 GFP感染的神经节神经元进行定量，用于证明特定T细胞亚群在防止神经节的急性和潜在HSV-2感染中的作用。 在第二个目标中，将使用抗体缺乏小鼠（MUMT）来确定HSV特异性抗体在保护感觉神经节中的作用。 HSV特异性血清的纯化IgG和IgA级分将用于HSV-免疫MUMT小鼠，以确定特异性抗体同型在完成保护HSV-rimmune Mumt小鼠保护潜在HSV-2感染的疗效。 在第三个目标中，将测试通过接种远端粘膜或全身部位保护感觉神经节的免疫反应的能力。 将评估这些位点的接种在阴道粘膜内引起长期记忆免疫反应的能力和相关的生殖器淋巴组织，可以迅速召回以保护阴道粘膜和感觉神经节。