In Vivo Imaging of Leukocyte-Endothelial Dynamics

描述（由申请人提供）：白细胞跨越血管内皮屏障的迁移过程是炎症和感染反应过程的基础。我们将量化各种药物的作用，如外用皮质类固醇、口服非甾体抗炎药和肥大细胞稳定剂在这些过程中的作用。我们已经应用活体显微镜在动物系统可视化，定量和分析这一过程。在共聚焦显微镜的最新进展已经允许一个欧洲小组定量白细胞内皮滚动和粘在微血管的人的眼睛。结合我们在共聚焦显微镜方面的临床专业知识和我们分析白细胞血管相互作用的经验，我们建议利用体内共聚焦技术来定量白细胞在4种不同的人体血管床上的滚动和停止：边缘、结膜、外巩膜和巩膜。有了这三个具体的目的：目的一，我们建议成像白细胞滚动和粘附在四个不同的正常眼部血管床：结膜，边缘，外巩膜和巩膜。目的二，我们将比较七种疾病状态下特定血管床的白细胞内皮动力学：a)过敏性季节性结膜炎，b)干燥综合征和干眼症，c)眼睑炎，d)移植物抗宿主病（GVHD）， e)巩膜炎，f)巩膜炎，g)前葡萄膜炎。目标三，我们将确定包括外用强的松龙醋酸酯、肥大细胞稳定剂（奥替萘洛尔）或口服非甾体抗炎药（吲哚美辛）在内的药物对经常开处方的疾病的内皮-白细胞动力学的影响。我们的研究将直接阐明几个棘手的和很少被研究的眼部疾病过程的发病机制。这些研究将阐明药物改变这些过程的机制。最重要的是，这些研究将量化微血管床的基本人类生物过程，这是以前没有成像的。

DESCRIPTION (provided by applicant): The process of leukocyte migration across the vascular endothelial barrier is fundamental to the process of inflammation and the response to infection. We will quantitate the effect of various medications such as topical corticosteroids, oral nonsteroidal anti-inflammatory drugs, and mast cell stabilizers in several of these processes. We have applied intravital microscopy in animal systems to visualize, quantitate and analyze this process. Recent advances in confocal microscopy have allowed a European group to quantitate leukocyte endothelial rolling and sticking in the microvasculature of the human eye. Combining our clinical expertise in confocal microscopy and our experience analyzing leukocyte vascular interactions, we propose to utilize in vivo confocal technology to quantitate leukocyte rolling and arrest in 4 different human vascular beds: the limbus, conjuctiva, episclera, and sclera. With these three specific aims: Aim one, we propose to image rolling and sticking of leukocytes in four different normal ocular vascular beds: the conjunctiva, limbus, episclera, and sclera. Aim two, we will compare leukocyte-endothelial dynamics in specific vascular beds in seven disease states: a) allergic seasonal conjunctivitis, b) Sjogren's syndrome and dry eye, c) blepharitis, d) graft versus host disease (GVHD), e) episcleritis, f) scleritis, and g) anterior uveitis. Aim three, we will determine the effect of medications including topical prednisolone acetate, a mast cell stabilizer (optipranolol), or an oral nonsteroidal anti-inflammatory drug (indomethacin) on endothelial-leukocyte dynamics in diseases for which each is frequently prescribed. Our studies will directly clarify the pathogenesis of several troublesome and rarely studied ocular disease processes. These studies will elucidate the mechanism by which medications alter these processes. Most importantly these studies will quantitate a fundamental human biological process in microvascular beds that have not previously been imaged.