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

描述（由申请人提供）：克莉丝汀约翰斯顿博士于2007年在华盛顿大学完成了传染病奖学金和流行病学硕士学位。该提案描述了一个为期5年的培训计划，这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。安娜沃尔德和劳伦斯科里博士是国际知名的领导人在临床HSV-2研究，并共同导师为这个奖项。 HSV-2是全球最流行的性传播感染，已成为获得和传播HIV的重要风险因素。HSV-2传播最常发生在无症状的生殖器脱落期间。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事件持续不到12小时，这表明神经节的频繁再激活率和活跃的外周免疫反应。初步数据表明，亚临床脱落可能同时发生在几个生殖器部位，HSV-2复制的“短爆发”发生在抑制性抗病毒治疗的存在下。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复制至关重要。为了评估“短爆发”的脱落，50名接受抑制性抗病毒治疗的HSV-2血清阳性患者的队列将获得生殖器拭子，每天4次，持续60天。将HSV-2排毒发作的发生率和持续时间与停止治疗的发生率进行比较（目标1）。对于目的2和3，将在30天内每天观察15名未接受治疗的HSV-2血清阳性者（目的2）和15名接受抑制性治疗的人（目的3），以使用新的快速HSV PCR测试捕获无症状脱落的发作。生殖器活检将在HSV-2检测后数小时内进行。将使用微阵列和HSV特异性T细胞功能评价基因表达模式。结果将与从存在生殖器病变的相同参与者获得的活检进行比较。这些研究将帮助我们了解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流行传播的治疗策略提供信息。

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.