Dynamics of the Ocular Immune Response During Uveitis

摘要 葡萄膜炎是一种潜在的致盲性疾病，是美国人失明的主要原因。特性 自身免疫性疾病的特征是靶组织炎症的复发和缓解。在自身免疫性疾病中 葡萄膜炎，眼睛是目标，并且因为眼睛允许光穿透大部分器官，所以对葡萄膜炎的研究 在眼睛的免疫反应中具有特别的优势。这个优点是光可以作为一种非侵入性的 一种标记炎症过程中存在的细胞并在炎症消退时迁移出来的方法。实验 自身免疫性葡萄膜炎（EAU）是最广泛使用的自身免疫性葡萄膜炎模型。我们建议生成并验证一个 稳定表达光诱导表达系统的新型小鼠，所述光诱导表达系统可用于在任何 在EAU（具体目标1）之前或期间。我们的假设是，一个转基因小鼠，表达光活化 Cre将是永久标记眼睛细胞的绝佳工具。目前，唯一可用的长期标签系统 依赖于注射可能有毒并扩散出靶组织的化合物。该提案利用了一种新颖的 驱动Cre活化以诱导允许永久标记的永久遗传变化的光活化系统 的靶细胞。第二代系统已在哺乳动物细胞系和小鼠中得到证实， 分离靶细胞。在这个提议中，我们将产生一种新的小鼠品系，其稳定表达用于 目的是回答与一个容易让光线通过的器官有关的免疫学问题。 重要的是，这将允许许多未来的研究，可以回答通过非侵入性标记的细胞在任何 任何组织中允许光穿透的时间。

Abstract Uveitis is a potentially blinding disease’s that are a leading cause of blindness among Americans. Characteristic of autoimmune diseases is marked relapsing and remitting inflammation of the target tissue. In the case of autoimmune uveitis, the eye is the target and because the eye allows for the penetration of light through much of the organ, study of the immune response in the eye has a particular advantage. This advantage being that light could be used as a non-invasive method to label cells that are present during inflammation and migrate out as the inflammation resolves. Experimental autoimmune uveitis (EAU) is the most widely used model of autoimmune uveitis. We propose to generate and validate a novel mouse that stably expresses a light inducible expression system that can be used to label cells in the eye during any point before or during EAU (Specific Aim 1). Our hypothesis is that a transgenic mouse that expresses photoactivatable Cre will be an excellent tool to permanently label cells in the eye. Currently, the only long-term labeling systems available rely on the injection of compounds that can be toxic and diffuse out of the target tissue. This proposal utilizes a novel photoactivatable system that drives Cre activation to induce permanent genetic changes that allow for permanent labeling of the target cells. A second-generation system has been demonstrated in mammalian cell lines, and in mice by transfecting the target cells. In this proposal we will generate a novel mouse line that stably expresses the transgenes for the purpose of answering the immunological questions that relate to an organ that readily allows for light to pass through. Importantly, this will allow for many future studies that can be answered through a non-invasive labeling of cells at any time in any tissue that allows for the penetration of light.