Therapeutic Ocular HSV Vaccine in HLA Transgenic Rabbits

单纯疱疹病毒1型（HSV-1）感染角膜，然后在感觉神经元中建立潜伏期， 三叉神经节（TG）。HSV-1的散发性自发再活化导致病毒在泪液中脱落 导致病毒传播给其他个体，也可引起复发性疱疹性基质角膜炎（HSK）， 致盲性眼病我们目前知识的一个主要空白是："我们如何预防或显著减少 泪液中的病毒脱落和HSV诱导的眼部疾病是由于潜伏病毒在泪液中的自发再活化引起的。 TG？"HSV特异性CD8 + T细胞在体外诱导的HSV-1再活化中似乎减少了 tg等不幸的是，小鼠中HSV-1的自发再活化是极其罕见的，因此这些研究的相关性是不确定的。 在小鼠中不能确定体内HSV-1自发再活化的发现。我们现在有一个"人性化"的 眼部HSV-1的HLA转基因兔模型，其产生"人样" CD8 T细胞免疫应答（HLA Tg兔）。在一项初步研究中，我们发现，治疗性免疫潜伏感染的HLA Tg兔， 来自HSV-1 gD的3个人CD8 T细胞表位使自发再活化降低4倍。这本小说 动物模型现在将允许我们第一次测试一种治疗性疫苗， 适当的人T细胞对HSV-1的应答可以降低自发再活化（病毒 眼睛脱落和HSV诱导的眼部疾病）。我们的具体目标包括： （一）.检验用HSV-1人CD8 + T细胞表位治疗性免疫可 减少潜伏感染的HLA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CD 4+和CD 8 + T细胞表位的不同组合（gB & gD）， 将用于免疫潜伏感染的HLA Tg兔。防止眼部病毒脱落（由于 自发再活化）和HSV诱导的眼部疾病。 （二）、验证HLA治疗性疫苗接种诱导的保护性免疫 目的1中的转基因兔与TG中效应和记忆CD8 + T细胞的存在相关， 结膜和/或引流淋巴结。我们将评估HSV-和表位的数量/功能是否与HSV-和表位的数量/功能相关。 体内诱导的特异性CD8 + T细胞与防止泪液中自发性病毒脱落相关， 眼部疾病我们将评估与保护相关的CD8 + T细胞机制。 （三）、检验潜伏感染HLA Tg兔中CD8 + T细胞减少将消除 疫苗效力，并增加未接种疫苗的家兔的自发再激活。 这些研究将提供关于CD8 + T细胞特异性作用的重要新信息， 人抗原表位在HSV-1自发再活化免疫控制中的作用这可能会导致 眼疱疹免疫策略的新范例。

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.