S-ANTIGEN--STRUCTURE/FUNCTION RELATIONSHIPS IN UVEITIS

S-抗原——葡萄膜炎中的结构/功能关系

• 批准号: 3259877
• 负责人: LARRY A DONOSO
• 金额: $ 8.59万
• 依托单位: WILLS EYE HEALTH SYSTEM
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-09-30 至 1987-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3259877
• 关键词: affinity chromatography; antigens; autoimmune disorder; chemical structure function; disease /disorder model; enzyme linked immunosorbent assay; gel filtration chromatography; high performance liquid chromatography; hybridomas; monoclonal antibody; spectrometry; tissue /cell culture; uveitis

In 1973 Kalsow and Wacker localized the antigen responsible for the induction of experimental autoimmune uveitis (EAU) in animals to completely surround the photoreceptor cells of the retina. Subsequently Wacker and Donoso (1975) isolated, purified and characterized this highly uveitopathogenic protein designated S-antigen from bovine retina. More recently S-antigen has been implicated in certain forms of human uveitis such as birdshot retinochoroidopathy and sympathetic ophthalmia. Recently Jakobiec and associates (1983) in an elegant study characterized sympathetic ophthalmia as a localized T cell delayed hypersensitivity abnormality. Despite numerous reports concerning the pathologic and immunologic feature of EAU, almost no information is available regarding the nature of the antigenic sites and structural/functional relationships of this important protein. We propose to study structure/function relationships of bovine and human retinal S-antigen. Biochemical studies will consist of producing slight modification of the intact native S-antigen. These modifications will be produced by using reagents which react with specific amino acid groups under mild conditions and which can be measured quantitatively. The effects of these modifications on the antigenicity of S-antigen will be determined by the ELISA assay and by using the animal model of EAU. The amino acid sequence of S-antigen will be determined using standard techniques and the automated amino acid sequenator. Immunologic studies will consist of the production of hybridoma secreted monoclonal antibodies by fusing spleen cells form mice immunized with S-antigen with the mouse myeloma cell line (SP 2/0.Ag 14). These antibodies will be utilized to examine differences in antigenic epitopes between bovine and human S-antigen These studies will lead to a better understanding of the biochemistry and immunology of S-antigen and may identify sites of immune interaction in EAU.

1973年，Kalsow和Wacker定位了导致这种病毒的抗原 实验性自身免疫性葡萄膜炎(EAU)动物模型的建立 包围视网膜的感光细胞。随后，瓦克和 Donoso(1975)对其进行了分离、纯化和高度鉴定 牛视网膜葡萄膜致病蛋白命名为S抗原。更多 最近，S抗原被认为与某些形式的人类葡萄膜炎有关 如鸟射视网膜脉络膜病和交感性眼炎。最近 Jakobiec and Associates(1983)在一项优雅的研究中 交感性眼炎是一种局限性T细胞迟发性超敏反应 反常现象。尽管有很多关于病理和病理的报道 EAU的免疫学特征，几乎没有关于EAU的信息 抗原性位点的性质和结构/功能关系 这种重要的蛋白质。 我们建议研究牛和人的结构/功能关系 视网膜S抗原。生化研究将包括产生轻微的 完整的天然S抗原的修饰。这些修改将是 使用与特定氨基酸基团发生反应的试剂生产的 在温和的条件下，可以定量测量。这个 这些修饰对S抗原抗原性的影响将是 用ELISA法和EAU动物模型测定。这个 S抗原的氨基酸序列将用标准确定 技术和氨基酸自动测序仪。免疫学研究 将由产生杂交瘤分泌的单抗组成 S抗原免疫的小鼠脾细胞与小鼠的融合 骨髓瘤细胞系(SP2/0.Ag14)。这些抗体将被用来 牛与人S抗原抗原表位差异的检测 这些研究将有助于更好地理解生物化学和 S抗原的免疫学，并可能识别EAU中的免疫相互作用部位。