Therapeutic Ocular HSV Vaccine in HLA Transgenic Rabbits

单纯疱疹病毒1型(HSV-1)感染角膜，然后在感觉神经元上建立潜伏期 三叉神经节(TG)。HSV-1的零星自发再激活导致泪液中的病毒脱落 导致病毒传播给其他人，还可能导致复发的疱疹间质性角膜炎(HSK)， 致盲的眼病。我们目前知识中的一个主要差距是：我们如何才能防止或显著减少 泪液中的病毒脱落和HSV引起的眼部疾病 HSV特异性CD8+T细胞在体外诱导的移植小鼠HSV-1再激活中的作用 Tg.不幸的是，在小鼠中，HSV-1的自发重新激活是极其罕见的，所以这些相关性 无法确定HSV-1在小鼠体内自发重新激活的结果。我们现在有了一个“人性化的” 构建类人CD8 T细胞免疫应答的眼部HSV-1转基因兔模型 TG兔)。在初步研究中，我们发现潜伏感染的HLATG兔的治疗性免疫 用HSV-1的3个人CD8T细胞表位，gD的自发再激活减少了4倍。这部小说 动物模型现在将首次允许我们检验这样的假设：一种治疗性疫苗可以诱导 适当的人类T细胞对HSV-1的反应可以减少自发再激活(病毒)的影响 眼睛脱落和单纯疱疹病毒引起的眼部疾病)。我们的具体目标包括： (1)。验证HSV-1人CD8+T细胞表位治疗性免疫可以 减少潜伏感染人类白细胞抗原转基因兔的自发再激活。CD4-CD8脂肽疫苗， 携带来自糖蛋白B和D的人类CD4+和CD8+T细胞表位的不同组合(GB和GD)， 将用于免疫潜伏感染的HLATG兔。防止眼睛中的病毒脱落(由于 自发性再激活)和单纯疱疹病毒引起的眼部疾病将被确定。 (2)。验证人类白细胞抗原治疗性疫苗诱导保护性免疫的假说 AIM 1中的转基因兔与TG中存在效应性和记忆性CD8+T细胞相关， 结膜和/或引流淋巴结节。我们将评估HSV-和表位的数量/功能- 体内诱导的特异性CD8+T细胞与保护泪液和泪液中自发的病毒脱落有关 眼疾。我们将评估与保护相关的CD8+T细胞机制。 (3)。验证潜伏感染的HLATG兔CD8+T细胞减少将被消灭的假设 疫苗的效力，也增加了未接种疫苗的兔子的自发再激活。 这些研究将提供有关CD8+T细胞所起作用的重要新信息 人类表位在HSV-1自发再激活免疫控制中的作用。这可能会导致 针对眼部疱疹的免疫治疗策略的新范例。

Herpes simplex virus 1 (HSV-1) infects the cornea and then establishes latency in sensory neurons of the trigeminal ganglia (TG). Sporadic spontaneous reactivation of HSV-1 causes shedding of virus in tears leading to spread of virus to other individuals, and can also cause recurrent Herpes Stromal Keratitis (HSK), a blinding ocular disease. A major gap in our current knowledge is: "How can we prevent or significantly reduce virus shedding in tears and HSV-induced ocular disease due to spontaneous reactivation of latent virus in the TG?" HSV-specific CD8+ T-cells appear to decrease in vitro induced HSV-1 reactivation in explanted mouse TG. Unfortunately, spontaneous reactivation of HSV-1 in mice is extremely rare so the relevance of these findings to in vivo HSV-1 spontaneous reactivation cannot be determined in mice. We now have a "humanized" HLA transgenic rabbit model of ocular HSV-1 that mounts "human-like" CD8 T-cell immune responses (HLA Tg rabbits). In a Preliminary Study we found that therapeutic immunization of latently infected HLA Tg rabbits with 3 human CD8 T-cell epitopes from HSV-1 gD decreased spontaneous reactivation 4-fold. This novel animal model will now allow us for the first time to test the hypothesis that a therapeutic vaccine that induces appropriate human T-cell responses to HSV-1 can decrease the effects of spontaneous reactivation (virus shedding in eyes and HSV-induced ocular disease). Our specific Aims include: (1). Test the hypothesis that therapeutic immunization with HSV-1 human CD8+ T-cell epitopes can decrease spontaneous reactivation in latently infected HLA Transgenic rabbits. CD4-CD8 lipopeptide vaccines, bearing different combinations of human CD4+ and CD8+ T cell epitopes from glycoprotein B and D (gB & gD), will be used to immunize latently infected HLA Tg rabbits. Protection against virus shedding in eyes (due to spontaneous reactivation) and HSV-induced ocular disease will be determined. (2). Test the hypothesis that the protective immunity induced by the therapeutic vaccination of HLA Transgenic rabbits in Aim 1 correlates with the presence of effector and memory CD8+ T-cells in the TG, conjunctiva, and/or draining lymph nodes. We will assess whether the number/function of HSV- and epitope- specific CD8+ T cells induced in vivo correlates with protection from spontaneous virus shedding in tears and ocular disease. We will assess the CD8+ T cell mechanism that correlates with protection. (3). Test the hypothesis that decreasing CD8+ T cells in latently infected HLA Tg rabbits will abrogate vaccine efficacy and also increase spontaneous reactivation in unvaccinated rabbits. These studies will provide important new information regarding the role of CD8+ T cells specific to human epitopes in immune control of HSV-1 spontaneous reactivation. This may lead to the development of new paradigms for immunotherapeutic strategies against ocular herpes.