PROTECTION OF GENITAL MUCOSA AND GANGLIA AGAINST HSV-2

大于单纯疱疹病毒2型（HSV-2）现在感染大约五分之一的美国人的生殖道。预防新的HSV-2感染的策略因其在感觉神经节中启动潜伏感染、周期性再激活并在生殖器组织中引起复发性病变或无症状病毒脱落从而增加其感染新宿主的机会的能力而变得复杂。 需要有效的疫苗来防止感觉神经节内潜伏期的建立。 不幸的是，我们对保护感觉神经节的免疫机制知之甚少。 在动物模型中，先前生殖器接种HSV-2可增强免疫力，保护感觉神经节免受再感染，因此可作为有效HSV-2疫苗的范例。 该提案的长期目标是使用生殖器HSV-2接种的小鼠模型来了解负责保护的免疫机制的类型，这些机制如何在分子水平上工作，以及如何引发这些反应以提供长期保护。 这些研究结果将为合理设计预防HSV-2的疫苗提供重要信息。在第一个目标中，表达绿色荧光蛋白（HSV-2 gfp）的重组HSV-2菌株将被用作标记物，以确定HSV特异性T淋巴细胞是否阻止HSV-2到达感觉神经节，从而防止潜伏期的建立。 通过UV显微镜对HSV-2 gfp感染的神经节神经元的定量和通过定量PCR对神经节中HSV-2 gfp基因组的定量将用于证明特异性T细胞亚群在预防神经节的急性和潜伏HSV-2感染中的作用。 在第二个目标中，将使用抗体缺陷型小鼠品系（muMT）来确定HSV特异性抗体在保护感觉神经节中的作用。 将HSV特异性血清的纯化IgG和伊加级分给予HSV免疫muMT小鼠，以确定特异性抗体同种型在完成HSV免疫muMT小鼠针对潜伏HSV-2感染建立的保护中的功效。 在第三个目标中，将测试通过接种远端粘膜或全身部位引起的免疫应答保护感觉神经节的能力。 将评估在这些部位接种以在阴道粘膜和相关生殖器淋巴组织内引发长期记忆免疫应答的能力，所述免疫应答可被快速召回以保护阴道粘膜和感觉神经节。

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.