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: 2757770
• 负责人: Gregg N. Milligan
• 金额: $ 17.09万
• 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 2004-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2757770
• 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.

目前，大约五分之一的美国人的生殖道受到 2 型单纯疱疹病毒 (HSV-2) 的感染。预防新的 HSV-2 感染的策略变得复杂，因为它能够在感觉神经节中启动潜伏感染，定期重新激活，并在生殖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。 需要有效的疫苗来防止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，先前生殖器接种 HSV-2 会引发免疫力，保护感觉神经节免遭再次感染，从而成为有效 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。 这些研究的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，从而阻止建立延迟。 通过UV显微镜对HSV-2 gfp感染的神经节神经元进行定量，并通过定量PCR对神经节中的HSV-2 gfp基因组进行定量，将用于证明特定T细胞亚群在预防神经节急性和潜伏HSV-2感染中的作用。 第二个目标是使用抗体缺陷小鼠品系 (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 IgA 级分给予 HSV 免疫 muMT 小鼠，以确定特异性抗体同种型在完成保护 HSV 免疫 muMT 小鼠免受潜伏 HSV-2 感染建立方面的功效。 在第三个目标中，将测试通过远端粘膜或全身部位的接种引起的免疫反应保护感觉神经节的能力。 将评估在这些部位的接种在阴道粘膜和相关生殖器淋巴组织内引发长期记忆免疫反应的能力，这些免疫反应可以被快速召回以保护阴道粘膜和感觉神经节。