Department of Ophthalmology, University of Washington.
Academic Unit of Ophthalmology, Translational Health Sciences, University of Bristol.
J Vis Exp. 2021 Dec 17(178). doi: 10.3791/62925.
The term 'uveitis' describes a heterogeneous set of conditions that all feature intraocular inflammation. Broadly, uveitis is defined by etiology: infection or autoimmunity. Infectious uveitis requires treatment with the appropriate antimicrobial agents, while autoimmune uveitis requires treatment with corticosteroids or other immunosuppressive agents. Post-infectious uveitis is a form of chronic uveitis that requires corticosteroids to control immune sequela following the initial infection. Uveitis associated with Mycobacterium tuberculosis (Mtb) infection is a well-recognized form of post-infectious uveitis, but the mechanisms of disease are not fully understood. To understand the role mycobacterial antigens and innate ligands play in stimulating chronic ocular inflammation following mTB infection, the model Primed Mycobacterial Uveitis (PMU) was developed for use in mice. This manuscript outlines the methods for generating PMU and monitoring the clinical course of inflammation using color fundus and optical coherence tomography (OCT) imaging. PMU is induced by immunization with heat-killed mycobacterial extract followed by intravitreal injection of the same extract into one eye seven days later. Ocular inflammation is monitored longitudinally using in vivo imaging and followed by sample collection for a wide range of assays, including histology, flow cytometry, cytokine analysis, qPCR, or mRNA sequencing. The mouse model of PMU is a useful new tool for studying the ocular responses to mTB, the mechanism of chronic uveitis, and for preclinical effectiveness tests of new anti-inflammatory therapies.
“葡萄膜炎”一词描述了一组具有不同特征的疾病，这些疾病都表现为眼内炎症。广义上，葡萄膜炎是根据病因来定义的：感染或自身免疫。感染性葡萄膜炎需要使用适当的抗菌药物进行治疗，而自身免疫性葡萄膜炎则需要使用皮质类固醇或其他免疫抑制剂进行治疗。感染后葡萄膜炎是一种慢性葡萄膜炎，需要使用皮质类固醇来控制初始感染后的免疫后遗症。与结核分枝杆菌（Mtb）感染相关的葡萄膜炎是一种公认的感染后葡萄膜炎形式，但疾病的机制尚未完全清楚。为了了解分枝杆菌抗原和天然配体在 mTB 感染后刺激慢性眼部炎症中的作用，开发了用于小鼠的 Primed Mycobacterial Uveitis (PMU) 模型。本文概述了使用热灭活分枝杆菌提取物进行免疫接种，然后在 7 天后将相同提取物眼内注射诱导 PMU，并使用眼底彩色照相和光学相干断层扫描（OCT）成像监测炎症的临床过程。PMU 通过免疫接种热灭活分枝杆菌提取物诱导，并在 7 天后将相同提取物眼内注射来诱导。通过体内成像进行纵向监测眼内炎症，并进行广泛的检测，包括组织学、流式细胞术、细胞因子分析、qPCR 或 mRNA 测序，以收集样本。PMU 小鼠模型是研究 mTB 对眼部的反应、慢性葡萄膜炎的机制以及新的抗炎治疗方法的临床前疗效的有用新工具。
