In Vivo Imaging of Leukocyte-Endothelial Dynamics

描述(申请人提供)：白细胞穿过血管内皮细胞屏障的过程是炎症过程和对感染反应的基础。我们将量化不同药物的效果，如外用皮质类固醇、口服非类固醇抗炎药和肥大细胞稳定剂。我们已经在动物系统中应用了活体显微镜来可视化、量化和分析这一过程。共聚焦显微镜的最新进展使一个欧洲研究小组能够量化白细胞内皮细胞在人眼微血管系统中的滚动和粘连。结合我们在共聚焦显微镜方面的临床专业知识和我们对白细胞血管相互作用的分析经验，我们建议利用体内共聚焦技术来量化4种不同的人类血管床中的白细胞滚动和停止：角膜缘、结膜、巩膜上膜和巩膜。有了这三个特定的目标：目标一，我们提出了四种不同的正常眼血管床：结膜、角膜缘、巩膜上膜和巩膜中白细胞的滚动和粘连的图像。目的比较过敏性季节性结膜炎、干燥综合征和干眼症、眼睑炎、移植物抗宿主病(GVHD)、硬膜炎、硬膜炎、前葡萄膜炎等七种疾病在特定血管床中的白细胞-内皮动力学变化。目的第三，我们将确定包括外用醋酸泼尼松龙、肥大细胞稳定剂(奥替普尼洛尔)或口服非类固醇抗炎药(吲哚美辛)在内的药物对经常使用这三种药物的疾病的内皮-白细胞动力学的影响。我们的研究将直接阐明几个棘手且很少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，这些研究将量化人类在微血管床上的一个基本生物过程，这些过程以前没有被成像过。

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.