ORIGIN OF ANTI-RO/SSA ANTIBODY

抗 RO/SSA 抗体的来源

• 批准号: 3085653
• 负责人: Robert Hal Scofield
• 金额: $ 7.67万
• 依托单位: OKLAHOMA MEDICAL RESEARCH FOUNDATION
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-03-15 至 1997-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3085653
• 关键词: Sjogren's syndrome; Vesiculovirus; antiantibody; antibody neutralization test; antigen antibody reaction; autoantibody; autoantigens; autoimmune disorder; chemical binding; human tissue; immunopathology; messenger RNA; northern blottings; nucleic acid sequence; nucleocapsid; plaque assay; polymerase chain reaction; protein structure; serology /serodiagnosis; southern blotting; systemic lupus erythematosus; virus protein; western blottings

Systemic lupus erythematosus, Sjogren's syndrome, and congenital complete heart block are unified by the presence of autoantibodies binding to Ro/SSA in a variable but substantial proportion (20% to 8O%) of patients with these diagnoses. Ro/SSA is an RNA-protein complex composed of a 60 kD peptide noncovalently bound to one of four small uridine rich RNAs. The sponsor's laboratory is dedicated to understanding the features of this autoantibody-autoantigen system from clinical and autoimmune response relationships to basic structural and antigenic characterization. Overlapping octapeptides have identified a major epitope of 60 kD Ro/SSA which is bound by greater than 50% of anti-Ro/SSA positive SLE patients. This epitope has an unexpected 7 of 8 amino acid sequence identity with the nucleocapsid (N) protein of the vesicular stomatitis virus (VSV). More unexpectedly, RO/SSA and the VSV N protein sequences share a total of six regions of short sequence identity. A reference anti-Ro/SSA serum binds five of the six regions (p=0.001), while 11 additional sera bind four of these six (p=0.003). This is the first known example of possible multiple site molecular mimicry between an autoantigen and a foreign protein. The project is designed to evaluate the fine specificity of the anti-Ro/SSA response and test the hypothesis that this response is immunologically related to VSV N protein. Overlapping peptides of both RO/SSA and VSV N will be prepared. Then the relative binding of anti-Ro/SSA and control sera will be appraised. VSV and related rhabdoviral strains will be used in antibody neutralizing assays and in Western blot assays in an effort to identify the virus most closely related to the anti-Ro/SSA response. Messenger RNA from patients and normal donors will be evaluated for the presence of rhabdovirarl sequences by the polymerase chain reaction and Southern blotting or by the direct evaluation of mRNA in Northern blots. These experiments have the potential to develop independent evidence for the presence of rhabdovirus in patients with anti-Ro/SSA autoantibodies. This project has the potential to at least preliminarily identify a pathogenic stimulus for the anti-Ro/SSA antibody.

系统性红斑狼疮、干燥综合征和先天性 完全性心脏传导阻滞因自身抗体的存在而统一 以可变但相当大的比例（20％至8O％）与Ro/SSA结合 患有这些诊断的患者。 Ro/SSA 是一种 RNA-蛋白质复合物 由与四个小肽之一非共价结合的 60 kD 肽组成 富含尿苷的RNA。 赞助商的实验室致力于 了解这种自身抗体-自身抗原系统的特征 临床和自身免疫反应与基本结构和 抗原表征。 重叠的八肽已确定 60 kD Ro/SSA 的主要表位，与超过 50% 的 抗Ro/SSA阳性SLE患者。 这个表位有一个意想不到的 7 个 8 氨基酸序列与核衣壳 (N) 蛋白的同一性 水泡性口炎病毒（VSV）。 更出乎意料的是，RO/SSA 和 VSV N蛋白序列共有六个短序列区域 身份。 参考抗 Ro/SSA 血清结合六个区域中的五个 (p=0.001)，另外 11 种血清与这 6 种血清中的 4 种结合 (p=0.003)。 这是第一个已知的可能的多位点分子的例子 自身抗原和外来蛋白质之间的拟态。 该项目是 旨在评估抗 Ro/SSA 反应的精细特异性 并检验该反应与免疫学相关的假设 VSV N 蛋白。 RO/SSA 和 VSV N 的重叠肽将是 准备好了。 然后抗Ro/SSA和对照血清的相对结合 将会被评价。 VSV 和相关弹状病毒株将用于 抗体中和测定和蛋白质印迹测定，以努力 识别与抗 Ro/SSA 反应最密切相关的病毒。 来自患者和正常捐赠者的信使 RNA 将被评估 通过聚合酶链式反应检测弹状病毒序列的存在，以及 Southern 印迹或通过 Northern 印迹直接评估 mRNA。 这些实验有可能为以下问题提供独立证据： 具有抗 Ro/SSA 自身抗体的患者中存在弹状病毒。 该项目有可能至少初步确定 抗 Ro/SSA 抗体的致病刺激。