PROTECTION OF GENITAL MUCOSA AND GANGLIA AGAINST HSV-2

第二型单纯疱疹病毒(HSV-2)现在感染着大约五分之一的美国人的生殖道。预防新的HSV-2感染的策略很复杂，因为它能够在感觉神经节启动潜伏感染，定期重新激活，并在生殖器组织中导致复发的病变或无症状的病毒脱落，从而增加感染新宿主的机会。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，人们对保护感觉神经节的免疫机制知之甚少。在动物模型中，以前生殖器接种HSV-2会引起免疫，保护感觉神经节免受再次感染，从而成为有效的HSV-2疫苗的范例。这项建议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发挥作用，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。在第一个目的中，将使用表达绿色荧光蛋白的重组HSV-2毒株(HSV-2 GFP)作为标记，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。用紫外显微镜定量检测HSV-2GFP感染的神经节细胞，用定量聚合酶链式反应检测神经节中HSV-2GFP基因组，以证明特定的T细胞亚群在预防神经节急性和潜伏感染HSV-2中的作用。在第二个目的中，将使用抗体缺陷小鼠株(MUMT)来确定HSV特异性抗体在保护感觉神经节中的作用。将纯化的HSV特异性血清的Ig G和Ig A部分应用于HSV免疫MUMA小鼠，以确定特定抗体亚型对HSV免疫MUM小鼠对抗HSV-2潜伏感染的保护作用。在第三个目标中，将测试通过接种远端粘膜或系统部位来保护感觉神经节的免疫反应的能力。在这些部位接种疫苗在阴道粘膜和相关的生殖器淋巴组织中引发长期记忆免疫反应的能力将被评估，这些组织可以迅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.