Vaccine-elicited genital and neuronal T cell responses

描述(申请人提供)：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。对HSV-2感染的保护很大程度上依赖于强大的抗原特异性T细胞反应的发展。鉴于HSV-2在感染后不久就能扩散到感觉神经节，疫苗诱导的T细胞需要被激活，表达效应功能，并在初始感染后迅速出现在适当的感染部位，如果疫苗要防止或极大地减少感觉神经节中潜伏病毒感染的建立。因此，用有效的HSV疫苗免疫应该导致效应器记忆T细胞部署到生殖器上皮细胞，可能也部署到感觉神经节。在拟议的研究中，我们将重点研究HSV疫苗诱导保护性T细胞免疫。具体地说，我们将研究HSV特异性记忆T细胞部署到生殖器上皮和感觉神经节的要求。在目标1中，我们将使用ELISPOT分析来检测免疫后感觉神经节中的HSV特异性记忆T细胞，并测试感觉神经节中对HSV特异性T细胞记忆的需求，以保护该组织免受HSV-2感染。目前正在开发的大多数HSV疫苗预计会激发体液免疫反应，主要是CD4+T细胞。在目标2中，我们将特异性地去除HSV免疫小鼠的T细胞亚群，以确定CD4+T细胞是否足以提供参与保护阴道上皮和感觉神经节的快速T细胞介导的成分。使用克隆型特异性单抗检测体内抗原特异性T细胞，我们将在目标3中测试仿照目前HSV疫苗临床试验使用的疫苗接种策略是否会导致记忆细胞部署到生殖器上皮和感觉神经节。我们还将测试是否可以通过1)在生殖道附近的粘膜或皮肤部位接种疫苗，以及2)增加疫苗激发T细胞反应的幅度，来改善记忆T细胞在这些组织中的部署。在目标4中，我们将使用传统的空斑分析和定量PCR分析来检测生殖器上皮和感觉神经节中的病毒，以测试与系统注射相同疫苗所产生的保护相比，针对生殖道的HSV疫苗是否能够产生更大的T细胞介导的对生殖器上皮和感觉神经节的保护作用。

DESCRIPTION (provided by applicant): Herpes simplex virus type 2 (HSV-2) now infects the genital tracts of approximately one in five Americans. Protection against HSV-2 infection is very dependent on the development of vigorous antigen-specific T cell responses. Given the ability of HSV-2 to spread to the sensory ganglia soon after infection, vaccine-elicited T cells will need to be activated, express effector function, and be present at the appropriate sites of infection very rapidly after initial infection if vaccines are to prevent or greatly reduce the establishment of latent virus infection in the sensory ganglia. Therefore, immunization with an effective HSV vaccine should result in the deployment of effector memory T cells to the genital epithelium and perhaps to the sensory ganglia as well. In the proposed studies we will focus on the induction of protective T cell immunity by HSV vaccines. Specifically we will examine the requirements for deployment of HSV-specific memory T cells to the genital epithelium and sensory ganglia. In Aim 1 we will use ELISPOT analysis to detect HSV-specific memory T cells in the sensory ganglia following immunization and test the requirement for HSV-specific T cell memory in the sensory ganglia to protect this tissue against HSV-2 infection. Most HSV vaccines currently being developed are expected to elicit humoral immune responses and predominantly CD4+ T cells. In Aim 2 we will specifically deplete T cell subsets from HSV-immune mice to determine if CD4+ T cells are sufficient to provide the rapid T cell mediated component involved in protection of the vagina epithelium and sensory ganglia. Using a clonotype-specific monoclonal antibody to detect antigen-specific T cells in vivo, we will test in Aim 3 if vaccination strategies modeled after those used in current HSV vaccine clinical trials will result in deployment of memory cells to the genital epithelium and sensory ganglia. We will also test if deployment of memory T cells to these tissues can be improved by 1) immunization at mucosal or cutaneous sites near the genital tract and 2) increasing the magnitude of the vaccine elicited T cell response. In Aim 4 we will use conventional plaque assays and quantitative PCR assays to detect virus in the genital epithelia and sensory ganglia to test if targeting an HSV vaccine to the genital tract results in greater T cell mediated protection of the genital epithelia and sensory ganglia compared to the protection resulting from systemic delivery of the same vaccine.