INTERLEUKIN-1 IN HERPES STROMAL KERATITIS

将使用良好表征的鼠眼部感染模型来研究 HSV-1诱导眼部感染的机制。 拟 促炎细胞因子白细胞介素1（IL-1）在 病毒感染。 一方面，它促进眼部炎症。 另一方面，它增强了限制HSV-1复制的防御机制 传播开来 为了验证这一假设，我们进行了一系列实验， 将进行IL-1同种型。 第一，原位技术 将使用杂交和免疫组织学来鉴定哪些角膜 细胞在HSV-1感染后最初产生IL-1 α和IL-1 β。 将进行实验以确定角膜细胞是否表达 体内IL-1受体。 接下来，将IL-1同种型接种到 角膜来测试它们刺激细胞因子的能力， 促炎和抗病毒活性。 细胞因子mRNA将 通过定量PCR检测，同时检测蛋白质产物 采用ELISA 最后，针对IL-1同种型的单克隆抗体 将在角膜内注射，以测试如何中和 内源性产生的IL-1影响HSV-1感染的早期事件。 的 角膜中研究的早期事件将是HSV-1复制，朗格汉斯 细胞迁移和促炎细胞因子合成。 总体看 从这些研究中获得的信息将有助于我们的长期 目标，即制定新的战略，以（a）尽量减少 角膜炎性反应，和（B）控制 这种重要的眼部病原体

A well characterized murine ocular infection model will be used to study mechanisms involved in HSV-1 induced ocular infection. It is proposed that the proinflammatory cytokine interleukin 1 (IL-1) plays a duel role in virus infection. On the one hand it promotes ocular inflammation. On the other, it enhances defense mechanisms that limit HSV-1 replication and spread. To test this hypothesis a series of experiments focusing on the IL-1 isoforms will be conducted. First, the techniques of in situ hybridization and immunohistology will be used to identify which corneal cells initially produce IL-1alpha and IL-1beta after HSV-1 infection. Experiments will be conducted to determine whether corneal cells express IL-1 receptors in vivo. Next, the IL-1 isoforms will be inoculated into the cornea to test their capacity to stimulate cytokines with proinflammatory and anti-viral activities. Cytokine mRNA will be detected by quantitative PCR while the protein product will be detected by ELISA. Finally, monoclonal antibodies specific for the IL-1 isoforms will be injected intracorneally in order to test how neutralization of endogenously produced IL-1 affects early events in HSV-1 infection. The early events studied in the cornea will be HSV-1 replication, Langerhans cell migration, and proinflammatory cytokine synthesis. Overall, the information gained from these studies will contribute to our long term objectives, which are to develop new strategies for (a) minimizing the corneal inflammatory response, and (b) controlling the replication of this important ocular pathogen.