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自身抗体是自身免疫的常见特征 全身性狼疮红斑，Sjogren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 已经发现了免疫遗传学和实验室关联 自身抗体。 此外，有强有力的证据表明抗RO/SSA 抗体集中在某些的皮肤，肾脏和腮腺 因此，患者能够致病。 这个项目 程序将首先表征50KD的顺序自动座 RO/SSA蛋白。 初步数据表明了精细的特异性 患者之间有所不同。 确实，在某些患者中显然很少 牛和人类RO/SSA之间的差异足以破坏 抗原性，并提出了人类RO/SSA抗原的假设 是自身免疫原。 人类cDNA蛋白序列可从我们的 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。 牛和人类序列 将用于研究60 kD之间的抗原差异 牛和人类RO/SSA蛋白。 重叠的肽已经 合成于13 kd的人类RO/SSA的羧基终端及其之一 确定的表位。 一旦顺序表位是初步的 确定，将以最佳尺寸和氨基酸为特征 顺序。 确实，可以揭示结构比 那些在人类ro/ssa中的人。 完成此操作后，精美的特殊性将是 在全身性狼疮患者的现有血清中测试 红细胞表评估与RO/SSA特定表位的抗体 已知的临床，实验室和免疫遗传学关系 ro/ssa。 在以后的实验中，精心选择的牛 - 人类杂种 RO/SSA的分子将制备并评估其抗原性。 这 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 重要风湿病自动抗原的重要结构特征 但也定义其最佳抗原结构以及不同的 临床，免疫遗传和实验室发现与自身的发现相关 RO/SSA在此自身抗体的精细特异性水平上。