Dynamic Reactivation of Herpes Simplex Virus-2 and the Genital Mucosal Response

描述(由申请人提供)：克里斯汀·约翰斯顿博士于2007年在华盛顿大学完成传染病奖学金和公共卫生硕士学位。这项建议描述了一项为期5年的培训计划，该计划将使她能够在研究生殖器单纯疱疹病毒-2(HSV-2)感染的临床研究中发展独立的学术生涯，并将建立在先前研究宿主-病毒相互作用和粘膜表面疱疹病毒复制的经验基础上。安娜·沃尔德博士和劳伦斯·科里博士是国际知名的HSV-2临床研究领导者，他们是该奖项的共同导师。 单纯疱疹病毒2型是全世界最流行的性传播感染，已成为感染和传播艾滋病毒的重要风险因素。HSV-2传播最常发生在无症状生殖器脱落期间。我们小组最近使用密集采样(每天擦拭生殖器表面4次)来证明大多数生殖器脱落持续时间不到12小时，这表明神经节经常重新激活，外周免疫反应活跃。初步数据表明，亚临床脱皮可能同时发生在几个生殖器部位，并且在抑制性抗病毒治疗存在的情况下，HSV-2复制的“短时间爆发”发生。这些数据表明，1)我们可能高估了抗病毒治疗提供的抑制程度，2)生殖器粘膜反应对于控制HSV-2复制是必不可少的。为了评估生殖器脱落的“短期爆发”，一组接受抑制性抗病毒治疗的50名HSV-2血清阳性患者将在60天内每天4次采集生殖器拭子。HSV-2脱落率和持续时间将与停止治疗的比率(目标1)进行比较。对于目标2和目标3，使用一种新的快速单纯疱疹病毒聚合酶链式反应测试，每天将观察15名未接受治疗的HSV-2阳性患者(目标2)和15名接受抑制治疗的患者(目标3)，以捕捉无症状脱皮的发作。生殖器活组织检查将在检测到HSV-2病毒后数小时内进行。使用微阵列的基因表达模式和HSV特异性T细胞功能将被评估。结果将与相同参与者在存在生殖器病变时进行的活组织检查进行比较。这些研究将有助于我们了解HSV-2的发病机制和介导病毒清除的反应，这将为预防HSV-2和HIV的流行传播提供治疗策略。

DESCRIPTION (provided by applicant): Dr. Christine Johnston completed Infectious Diseases fellowship and an MPH degree in epidemiology at the University of Washington in 2007. This proposal describes a 5 year training program which will allow her to develop an independent academic career in clinical research studying genital herpes simplex virus-2 (HSV-2) infection, and will build on prior experiences studying host-viral interactions and herpesvirus replication at mucosal surfaces. Drs. Anna Wald and Lawrence Corey are internationally renowned leaders in clinical HSV-2 research and are co-mentors for this award. HSV-2 is the most prevalent sexually transmitted infection worldwide and has emerged as a significant risk factor for acquisition and transmission of HIV. HSV-2 transmission most frequently occurs during periods of asymptomatic genital shedding. Our group has recently used intensive sampling (swabbing the genital surfaces 4 times per day) to demonstrate that the majority of genital shedding episodes last less than 12 hours, suggesting a frequent reactivation rate from the ganglia and a brisk peripheral immune response. Preliminary data indicate that subclinical shedding may occur simultaneously at several genital locations and that "short bursts" of HSV-2 replication occur in the presence of suppressive antiviral therapy. These data suggest that 1) we may have overestimated the degree of suppression provided by antiviral therapy and 2) genital mucosal responses are essential for control of HSV-2 replication. To evaluate "short bursts" of shedding, a cohort of 50 HSV-2 seropositive persons on suppressive antiviral therapy will obtain genital swabs 4 times per day for 60 days. The rate and duration of HSV-2 shedding episodes will be compared with rates off therapy (Aim 1). For Aims 2 & 3,15 HSV-2 seropositive people not on therapy (Aim 2) and 15 persons on suppressive therapy (Aim 3) will be seen daily over a 30 day period to capture episodes of asymptomatic shedding, using a novel rapid HSV PCR test. Genital biopsies will be performed within hours of HSV-2 detection. Gene expression patterns using microarrays and HSV specific T-cell function will be evaluated. Results will be compared to biopsies obtained from the same participants in the presence of genital lesions. These studies will help us understand HSV-2 pathogenesis and the responses mediating viral clearance, which will inform therapeutic strategies to prevent the epidemic spread of HSV-2 and HIV.