Purification of an acidic nuclear protein antigen and demonstration of its antibodies in subsets of patients with sicca syndrome.
Purification of an acidic nuclear protein antigen and demonstration of its antibodies in subsets of patients with sicca syndrome.
复制标题
酸性核蛋白抗原的纯化及其抗体在干燥综合征患者亚群中的证明。
作者：
Masashi Akizuki;Marilyn Boehm;S. Kassan;Alfred D. Steinberg;T. Chused
Ha 是一种可溶性核蛋白，通过连续应用分级硫酸铵沉淀（60% 至 80% 饱和度）、DEAE-Sephadex 层析（在 0.05 M Tris HCl 缓冲液中的 0.26 至 0.38 M NaCl 之间洗脱，pH 7.2）以及利用与来自具有高滴度 Ha 抗体的患者的 IgG 偶联的 Sepharose 免疫吸收柱的亲和层析，从小牛胸腺核提取物中纯化 Ha。 Ha抗原可以通过Bolton和Hunter的方法进行碘化，但不能通过氯胺-T或乳过氧化物酶方法进行碘化，表明不存在酪氨酸和组氨酸。将碘化材料通过 Sephadex G-100 柱以除去少量污染物。最终产物在聚丙烯酰胺凝胶电泳中迁移为单条带。利用125I-Ha抗原通过硫酸铵法测定特异性抗体。在没有其他结缔组织疾病的情况下，73% 的干燥综合征患者和 85% 与系统性红斑狼疮相关的干燥综合征患者观察到血清结合能力升高。在与类风湿性关节炎相关的干燥综合征（6%）和不伴有干燥综合征的系统性红斑狼疮中（3%），这种情况并不常见。在患有其他结缔组织疾病的患者或正常对照中未发现抗-Ha 抗体。 Ha 抗体是一部分干燥综合征患者的特征。
Ha, a soluble nuclear protein, was purified from calf thymus nuclear extract by successive application of fractional ammonium sulfate precipitation (60 to 80% saturation), DEAE-Sephadex chromatography (eluted between 0.26 and 0.38 M NaCl in 0.05 M Tris HCl buffer, pH 7.2), and affinity chromatography utilizing an immunoabsorbent column of Sepharose coupled to IgG from a patient with a high titer of antibody to Ha. The Ha antigen could be iodinated by the method of Bolton and Hunter but not by the chloramine-T or lactoperoxidase methods indicating the absence of tyrosine and histidine. The iodinated material was passed through a Sephadex G-100 column to remove minor contaminants. The final product migrated as a single band in polyacrylamide gel electrophoresis. The 125I-Ha antigen was utilized to measure specific antibody by the ammonium sulfate method. Elevated serum-binding capacity was observed in 73% of patients with sicca syndrome in the absence of another connective tissue disease and 85% of patients with sicca syndrome associated with systemic lupus erythematosus. It was infrequent in sicca syndrome associated with rheumatoid arthritis (6%) and in systemic lupus erythematosus without sicca syndrome (3%). Anti-Ha antibodies were not found in patients with other connective tissue diseases or normal controls. Antibody to Ha characterizes a subset of patients with sicca syndrome.