Rischmueller M, McNeilage L J, McCluskey J, Gordon T
Centre for Transfusion Medicine and Clinical Immunology, Flinders Medical Centre, Bedford Park, South Australia.
Clin Exp Immunol. 1995 Jul;101(1):39-44. doi: 10.1111/j.1365-2249.1995.tb02274.x.
In systemic autoimmunity, the human B cell response to the La (SS-B) autoantigen is polyclonal and directed to both conserved and human-specific epitopes. This study has further characterized the B cell epitope(s) present within the conserved central region of the La protein, LaC (amino acids 111-242) containing the RNA recognition motif (RRM, aa 111-187). Ten overlapping and non-overlapping protein fragments spanning LaC were expressed in bacteria as NH2-terminal fusions with glutathione-S-transferase. The fusion proteins were tested by ELISA for reactivity with a panel of human anti-La sera in order to define the nature of the epitopes. Ninety-two percent of patient sera containing anti-La antibodies reacted with the region of La containing the RRM. Fine mapping of this reactivity using deletion mutants indicated that the deletion of 19 amino acids from either the NH2-terminal or COOH-terminal region of the RRM was associated with loss of antibody reactivity, suggesting that the immunodominant epitope expressed in this region is discontinuous. Autoantibodies affinity-purified from the La RRM fragment to remove other specificities immunoprecipitated newly synthesized native La (SS-B)/Ro (SS-A) complexes, providing additional evidence that autoantibodies were recognizing a conformational epitope. The findings indicate that the human autoantibody response to La involves recognition of a conformational determinant involving the conserved RRM region without necessarily interfering with the RNA-dependent association of the La/Ro ribonucleoprotein.
在系统性自身免疫中,人类B细胞对La(SS - B)自身抗原的反应是多克隆的,且针对保守表位和人类特异性表位。本研究进一步对La蛋白保守中心区域(LaC,氨基酸111 - 242)内存在的B细胞表位进行了表征,该区域包含RNA识别基序(RRM,氨基酸111 - 187)。跨越LaC的10个重叠和非重叠蛋白片段在细菌中作为与谷胱甘肽 - S - 转移酶的氨基末端融合蛋白表达。通过ELISA检测融合蛋白与一组人类抗La血清的反应性,以确定表位的性质。92%含有抗La抗体的患者血清与包含RRM的La区域发生反应。使用缺失突变体对这种反应性进行精细定位表明,从RRM的氨基末端或羧基末端区域缺失19个氨基酸与抗体反应性丧失相关,这表明该区域表达的免疫显性表位是不连续的。从La RRM片段亲和纯化以去除其他特异性的自身抗体免疫沉淀了新合成的天然La(SS - B)/Ro(SS - A)复合物,提供了额外证据表明自身抗体识别的是构象表位。这些发现表明,人类对La的自身抗体反应涉及对涉及保守RRM区域的构象决定簇的识别,而不一定干扰La/Ro核糖核蛋白的RNA依赖性结合。