Correlating HSV-2 Disease Severity with Tissue Resident CD8 T-cells

单纯疱疹病毒2型（HSV-2）感染是终身的，是生殖器溃疡疾病的主要原因 国际吧生殖器HSV-2再激活的临床结果差异很大，从完全无症状到 罕见和短暂复发频繁和严重的生殖器溃疡：这种变化的原因 不知道。使用人类志愿者的生殖器皮肤和粘膜活检，我们已经表明HSV-2再激活导致局部免疫应答的长期持续。CD 8 T细胞持续存在于 真皮-表皮连接处（DEJ），邻近释放病毒体的感觉神经末梢。的 独特的解剖学分布表明，局部CDS T细胞可能在快速遏制中发挥关键作用， 病毒感染的外周，从而影响HSV-2疾病的临床和病毒学过程中， 人类在上一个资助周期，我们开发了一种细胞类型特异性激光捕获显微切割（LCM） 原位分离单个CD 8 T细胞并测量其活性的方法。通过采用以下综合办法， CD 8特异性LCM、全基因组转录谱分析和TCR库深度测序，我们现在可以 阐明组织驻留记忆CD 8 T细胞与生殖器疱疹疾病严重程度之间的关系， 人类在这个项目中，我们将研究HSV-2疾病严重程度与人类HSV-2复发部位组织驻留记忆CD 8 T细胞的数量，质量和多样性之间的关系。 我们的具体目标是：1）确定组织驻留记忆CD 8 T细胞的解剖学分布、密度、衰变动力学和抗病毒特征基因在患有轻度与重度生殖器HSV-2疾病的参与者中是否不同; 2）确定组织驻留记忆CD 8 T细胞的T细胞受体（TCR）库动力学和抗原特异性是否与生殖器疱疹疾病的严重程度相关。我们将从具有不同疾病结局的患者队列中获得连续活检组织：轻度疾病，定义为< 2 episodes per year and severe disease as recurrence rate >每年6次复发率。该项目将定义对HSV-2的成功外周免疫应答的特征，这些特征可以在疫苗开发期间作为免疫的潜在相关性。

Herpes simplex virus type-2 (HSV-2) infections are lifelong, and a leading cause of genital ulcer disease worldwide. The clinical outcome of genital HSV-2 reactivation varies widely from completely asymptomatic to infrequent and short-lasting recurrences to frequent and severe genital ulcerations: reasons for this variability are not known. Using biopsies of genital skin and mucosa from human volunteers, we have shown that HSV-2 reactivation results in a long-term persistence of local immune responses. CD8 T cells persist at the dermal-epidermal junction (DEJ) contiguous to the sensory nerve endings where virions are released. The unique anatomical distribution suggests that local CDS T cells might play a pivotal role in rapid containment of viral infection in the periphery, thus influencing the clinical and virologic course of HSV-2 disease in humans. In the last grant cycle, we developed a cell-type specific laser capture microdissection (LCM) method to isolate individual CD8 T cells in situ and measure their activity. By using combined approaches of CD8-specific LCM, whole genome transcriptional profiling and TCR repertoire deep sequencing, we can now elucidate associations between tissue resident memory CD8 T cells and genital herpes disease severity in humans. In this project, we will investigate the association between HSV-2 disease severity and the quantity, quality and diversity of tissue resident memory CD8 T cells at the site of previous HSV-2 recurrence in humans. Our specific aims are: 1) to define whether the anatomic distribution, density, decay kinetics and the antiviral signature genes of tissue resident memory CD8 T cells differ in participants with mild versus severe genital HSV-2 diseases; 2) to define whether the T cell receptor (TCR) repertoire dynamics and antigenic specificity of tissue resident memory CD8 T cells are associated with genital herpes disease severity. We will obtain sequential biopsy tissues from patient cohorts with distinct disease outcomes: mild disease, defined as recurrence rate < 2 episodes per year and severe disease as recurrence rate > 6 episodes per year. This project will define the characteristics of a successful peripheral immune response to HSV-2 that can be harnessed as a potential correlate of immunity during vaccine development.