Protection of Genital Mucosa and Ganglia Against HSV-2

描述(申请人提供)：大约五分之一的美国人感染了单纯疱疹病毒2型(HSV-2)，生殖器单纯疱疹病毒病每年造成的直接损失大约超过2亿美元。除了单纯疱疹病毒感染对新生儿的破坏性影响外，由单纯疱疹病毒引起的生殖器损害是感染艾滋病毒的主要风险因素。对于许多免疫功能低下的患者来说，疾病复发的频率增加以及无法解决口腔和肛门HSV损害仍然是一个严重的并发症。目前还没有获得许可的预防HSV疾病的疫苗。最近在临床试验中测试的疫苗产生了高滴度的特异性抗体，但并不能保护所有人免受HSV-2感染。拟议的研究将集中于特定T细胞亚群的保护性反应，并检查它们在生殖器上皮、感觉神经节和脊髓中作用的机制，以解决HSV感染。 在目标1中，我们将HSV免疫的CD4+和CD8+T细胞群转移到骨髓嵌合小鼠，以检测上皮细胞、天然免疫细胞和干扰素-γ在清除生殖道HSV中的作用。我们将确定干扰素-γ的细胞靶点是造血细胞还是体细胞，并研究干扰素-γ介导的该部位保护的分子机制。HSV-1也正在成为一种重要的生殖器病原体。在目标2中，我们将确定负责预防生殖器HSV-1感染的T细胞亚群。我们将利用ELISPOT和四聚体染色来量化Recall T细胞的反应，并评估它们在异型生殖器HSV攻击后保护感觉神经节的能力。在目标3中，我们将分析CD4+T细胞在神经元组织中的保护作用。我们将使用细胞耗竭和过继转移实验来测试HSV特异性召回抗体反应和神经元CD8+T细胞反应对CD4+T细胞的依赖性。此外，我们将利用免疫CD4+T细胞过继转移到免疫缺陷小鼠来确定CD4+T细胞是否有助于HSV从神经元组织中的清除。这些研究的结果对于HSV疫苗的合理开发以及了解HSV病变解决所需的细胞相互作用和特定的分子机制应该是重要的。

DESCRIPTION (provided by applicant): Approximately 1 in 5 Americans are infected with herpes simplex virus type 2 (HSV-2) and the approximate direct cost of genital HSV disease exceeds $200 million annually. In addition to the devastating effects of HSV infections in newborns, genital lesions caused by HSV represent a major risk factor for acquisition of HIV. The increased frequency of recurrent disease and the inability to resolve oral and anogenital HSV lesions remains a serious complication for many immunocompromised patients. There is no licensed vaccine for the prevention of HSV disease. Vaccines recently tested in clinical trial elicited high titers of specific antibody but did not protect all populations against HSV-2 infection. The proposed studies will focus on the protective responses by specific T cell subsets and examine the mechanisms by which they act in the genital epithelium, sensory ganglia, and spinal cord to resolve HSV infections. In Aim 1, we will transfer HSV-immune CD4+ and CD8+ T cell populations to bone marrow chimeric mice to examine the role of epithelial cells, innate immune cells, and IFN-gamma in clearance of HSV from the genital tract. We will determine if the cellular targets for IFN-gamma are hemopoietic or somatic cells and examine the molecular mechanisms of IFN-gamma -mediated protection of this site. HSV-1 is also emerging as an important genital pathogen. In Aim 2, we will identify the T cell subsets responsible for protection against genital HSV-1 infections. We will utilize ELISPOT and Tetramer staining to quantify recall T cell responses and assess their ability to protect the sensory ganglia following heterotypic genital HSV challenge. In Aim 3, we will analyze the protective role of CD4+ T cells in neuronal tissue. We will use cell depletion and adoptive transfer experiments to test the dependence of the HSV-specific recall antibody response and neuronal CD8+ T cell response on CD4+ T cells. Additionally, we will utilize adoptive transfer of immune CD4+ T cells to immunodeficient mice to determine if CD4+ T cells contribute to HSV clearance from neuronal tissue. The results of these studies should be important for the rational development of HSV vaccines as well as understanding the cellular interactions and specific molecular mechanisms required for resolution of HSV lesions.