AUTOANTIGENICITY OF THE 60 KD RO/SSA PROTEIN

60 KD RO/SSA 蛋白的自身抗原性

• 批准号: 3803416
• 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/3803416
• 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自身抗体是自体免疫的共同特征 系统性红斑狼疮、干燥综合征、亚急性皮肤性狼疮 红斑狼疮和新生儿狼疮综合征。无数临床病例 已经发现免疫遗传学和实验室与此有关 自身抗体。此外，有强有力的证据表明，抗Ro/SSA 抗体集中在某些人的皮肤、肾脏和腮腺。 患者，因此，能够是致病的。这一项目是美国 程序将首先表征50kD的顺序自动表位 RO/SSA蛋白。初步数据表明，这种良好的特异性 因病人不同而有所不同。事实上，在一些患者中，明显的轻微症状 牛和人的Ro/SSA的差异足以摧毁 抗原性，并导致假设人类Ro/SSA抗原 是一种自身免疫原。人类cdna蛋白序列可从我们的 以前的工作。在这些实验中，60kD牛Ro/SSA的cDNA 将被鉴定、克隆和测序。牛和人的序列 将被用来研究60kD之间的抗原差异 牛和人的Ro/SSA蛋白。重叠的多肽已经被 人Ro/SSA羧基末端13kD的合成及其一种 确定了表位。一旦序列表位初步确定 对它们进行鉴定，确定它们的最佳大小和氨基酸 序列。事实上，可能会揭示出比这更活跃的结构。 人类RO/SSA中的那些。一旦做到了这一点，就会有很好的针对性 在现有的系统性红斑狼疮患者血清收集中进行检测 红斑狼疮评估Ro/SSA特定表位抗体的方法 已知的临床、实验室和免疫遗传学与抗-HBs的关系 RO/SSA。在后来的实验中，精心挑选的牛和人的杂交 将制备Ro/SSA分子并对其抗原性进行评估。这个 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 还可以确定其最佳的抗原结构以及不同的 与自身抗体相关的临床、免疫遗传学和实验室结果 RO/SSA达到该自身抗体良好的特异性水平。