Hou Aihua, Bose Tanima, Chandy K George, Tong Louis
Singapore Eye Research Insitute, A Member of SingHealth; DUKE-National University of Singapore Medical School.
Lee Kong Chian School of Medicine, Nanyang Technological University.
J Vis Exp. 2017 Jun 7(124):55592. doi: 10.3791/55592.
Dry eye disease is a very common condition that causes morbidity and healthcare burden and decreases the quality of life. There is a need for a suitable dry eye animal model to test novel therapeutics to treat autoimmune dry eye conditions. This protocol describes a chronic autoimmune dry eye rat model. Lewis rats were immunized with an emulsion containing lacrimal gland extract, ovalbumin, and complete Freund's adjuvant. A second immunization with the same antigens in incomplete Freund's adjuvant was administered two weeks later. These immunizations were administered subcutaneously at the base of the tail. To boost the immune response at the ocular surface and lacrimal glands, lacrimal gland extract and ovalbumin were injected into the forniceal subconjunctiva and lacrimal glands 6 weeks after the first immunization. The rats developed dry eye features, including reduced tear production, decreased tear stability, and increased corneal damage. Immune profiling by flow cytometry showed a preponderance of CD3 effector memory T cells in the eyeball.
干眼疾病是一种非常常见的病症,会导致发病和医疗负担,并降低生活质量。需要一种合适的干眼动物模型来测试用于治疗自身免疫性干眼病症的新型疗法。本方案描述了一种慢性自身免疫性干眼大鼠模型。用含有泪腺提取物、卵清蛋白和完全弗氏佐剂的乳剂对Lewis大鼠进行免疫。两周后,用不完全弗氏佐剂中的相同抗原进行第二次免疫。这些免疫接种在尾根部皮下进行。为增强眼表和泪腺的免疫反应,在首次免疫6周后,将泪腺提取物和卵清蛋白注射到穹窿结膜下和泪腺中。大鼠出现了干眼特征,包括泪液分泌减少、泪液稳定性降低和角膜损伤增加。通过流式细胞术进行的免疫分析显示眼球中CD3效应记忆T细胞占优势。