HSV 1 LATENCY--IMMUNIZATION AND RECURRENT OCULAR DISEASE

拟议研究的长期目标是降低 复发性疱疹所致角膜盲的发生率 角膜炎。为实现这一长期目标而提出的第一个具体目标 目的包括评估活体减重的可行性 通过检验疱疹病毒疫苗的原始定植假说 携带HSV-1 TK毒株的三叉神经节将有效 用HSV-1 TK+野生型毒株防止二次定植。 具体地说，我们将确定是否可以占据一个以上的HSV-1毒株 潜伏期相同的神经节，如果HSV-1 TK强毒株 阻断野生型TK+菌株的定植和/或重新激活，如果 强毒型HSV-1毒株可与AFP互补或重组 无毒免疫病毒株产生潜伏的强毒株 能够产生复发性眼病的。隐匿性三叉神经 三叉神经与三叉神经共培养可检出疱疹病毒 神经节和离子导入引起的眼球流泪。已恢复 潜伏病毒将以Tk表型为特征，并通过 限制性内切酶分析。物种变异的作用来解释 目前文献中相互矛盾的结果将在老鼠身上进行评估。 还有兔子。实现以下长期目标的第二个具体目标 减少复发性疱疹病毒性角膜炎的失明包括阐明 潜伏期神经节细胞复活过程中的初始分子事件 疱疹。对早期基因转录的基本了解，包括 重新激活信号及其控制，对于实现第二个是必不可少的 长期目标。具体地说，我们将研究早期病毒RNA 原位杂交在神经节细胞中的转录。这些研究 将在兔身上使用6-羟基多巴胺加局部用药 肾上腺素离子导入诱发潜伏性神经节细胞再激活模型 疱疹病毒。这两个具体目标的结果将导致更好的 了解疱疹病毒潜伏期，并评估其潜伏期 活病毒疫苗的效力。

The long term ojbective of the proposed study is to reduce the high incidence of corneal blindness resulting from recurrent herpetic keratitis. The first proposed specific aim to meet this long term objective includes evaluating the feasibility of a live, attenuated herpesvirus vaccine by testing the hypothesis that primary colonization of the trigeminal ganglia with an avirulent HSV-1 TK- strain will effectively prevent secondary colonization with wild-type HSV-1 TK+ strains. Specifically, we will determine if more than one HSV-1 strain can occupy the same ganglion during latency, if an avirulent HSV-1 TK-strain will block colonization and/or reactivation of wild-type TK+ strains, and if a virulent challenge HSV-1 strain can complement or recombine with an avirulent immunizing virus strain to produce a latent virulent strain capable of producing recurrent ocular disease. Latent trigeminal herpesvirus(es) will be detected by both co-cultivation of trigeminal ganglia and iontophoresis-induced ocular shedding in tears. Recovered latent viruses will be characterized by TK phenotype and identified by restriction enzyme analysis. The role of species variations to explain contradictory results in the current literature will be evaluated in mice and rabbits. The second specific aim to achieve the long term objective of reducing blindness from recurrent herpetic keratitis includes elucidating the initial molecular events during reactivation of latent ganglionic herpes. A basic understanding of early gene transcription, including the reactivation signal and its control, is essential to achieve the second long-term objective. Specifically, we will study early virus RNA transcription in ganglionic neurons by in situ hybridization. The studies will be carried out in rabbits using 6 hydroxydopamine plus topical epinephrine iontophoresis model to induce reactivation of latent ganglionic herpesvirus. The results of both specific aims will lead to a better understanding of herpesvirus latency, and to an evaluation of the potential efficacy of a live virus vaccine.