AUTOANTIGENICITY OF THE 60 KD RO/SSA PROTEIN

60 KD RO/SSA 蛋白的自身抗原性

• 批准号: 3809932
• 负责人: JOHN B HARLEY
• 金额: --
• 依托单位: OKLAHOMA MEDICAL RESEARCH FOUNDATION
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3809932
• 关键词: Sjogren's syndrome; antibody specificity; autoantibody; autoantigens; autoimmune disorder; conformation; epitope mapping; human tissue; immunoglobulin genes; molecular cloning; protein structure function; ribonucleoproteins; synthetic antigens; systemic lupus erythematosus

Anti-Ro/SSA autoantibodies are a common feature of the autoimmunity of systemic lupus erythematosus, Sjogren's syndrome, subacute cutaneous lupus erythematosus, and neonatal lupus syndrome. Numerous clinical immunogenetic and laboratory associations have been found with this autoantibody. Moreover, there is strong evidence that anti-Ro/SSA antibodies are concentrated in the skin, kidney, and parotid glands of some patients and, hence, are capable of being pathogenic. This project of the Program will first characterize the sequential autoepitopes of the 50kD Ro/SSA protein. The preliminary data demonstrate that the fine specificity varies between patients. Indeed, in some patients the apparently minor differences between bovine and human Ro/SSA are sufficient to destroy antigenicity and have led to the hypothesis that the human Ro/SSA antigen is an autoimmunogen. The human cDNA protein sequence is available from our previous work. In these experiments, the cDNA for the 60 kD bovine Ro/SSA will be identified, cloned and sequenced. The bovine and human sequence will be used to investigate the antigenic differences between the 60 kD bovine and human Ro/SSA proteins. Overlapping peptides have been synthesized to the carboxy terminal 13 kD of human Ro/SSA and one of its epitopes identified. Once sequential epitopes are preliminarily identified, they will be characterized for optimal size and amino acid sequence. Indeed, structures may be revealed that are more reactive than those in human Ro/SSA. Once this has been done, fine specificity will be tested in existing collections of sera from patients with systemic lupus erythematosus to assess antibodies to particular epitopes of Ro/SSA with the known clinical, laboratory and immunogenetic relationships to anti- Ro/SSA. In later experiments, carefully selected bovine-human hybrid molecules of Ro/SSA will be prepared and their antigenicity assessed. The reactivity of these molecules will lead toward an understanding of the conformational epitopes. This work has the potential not only to reveal important structural features of an important rheumatic disease autoantigen but also to define its optimal antigenic structure as well as the different clinical, immunogenetic and laboratory findings associated with autoanti- Ro/SSA at the level of the fine specificity of this autoantibody.

抗Ro/SSA自身抗体是免疫性自身免疫的共同特征。 系统性红斑狼疮，干燥综合征，亚急性皮肤狼疮 红斑和新生儿狼疮综合征。 许多临床 免疫遗传学和实验室协会已被发现与此有关， 自身抗体 此外，有强有力的证据表明， 抗体集中在皮肤、肾脏和腮腺中， 患者，因此能够是致病性的。 这个项目的 程序将首先表征50 kD的连续自身表位 Ro/SSA蛋白。 初步数据表明， 患者之间有所不同。 事实上，在一些患者中， 牛和人Ro/SSA之间的差异足以破坏 抗原性，并导致假设，人Ro/SSA抗原 是自身免疫原 人cDNA蛋白质序列可从我们的实验室获得。 以前的工作。 在这些实验中，60 kD牛Ro/SSA的cDNA 将被识别，克隆和测序 牛和人的基因序列 将用于研究60 kD之间的抗原差异 牛和人Ro/SSA蛋白。 重叠肽已经被 合成到人Ro/SSA的羧基末端13 kD，并且其 鉴定的表位。 一旦序列表位被初步确定， 鉴定后，它们将被表征为最佳大小和氨基酸 顺序 事实上，结构可能会被揭示，更反应性比 在人类Ro/SSA中。 一旦做到这一点，精细的特异性将是 在现有的系统性狼疮患者血清中进行了检测 评估Ro/SSA特定表位的抗体， 已知的临床、实验室和免疫遗传学与抗- Ro/SSA。 在后来的实验中，精心挑选的牛-人杂交 制备Ro/SSA分子并评估其抗原性。 的 这些分子的反应性将导致对 构象表位 这项工作不仅有可能揭示 一种重要风湿病自身抗原的重要结构特征 还可以确定其最佳抗原结构以及不同的 与自身抗- Ro/SSA在该自身抗体的精细特异性水平上。