Wang X H, Miyazaki Y, Shinomura Y, Moriyama Y, Nakamoto R K, Matsuzawa Y, Maeda M, Futai M
Department of Organic Chemistry and Biochemistry, Osaka University, Japan.
Biochem Biophys Res Commun. 1993 Jan 15;190(1):207-14. doi: 10.1006/bbrc.1993.1032.
Human sera from autoimmune gastritis patients containing autoantibodies to gastric parietal cells were analyzed by immunological methods. Enzyme linked immuno-sorbent assay demonstrated that all nine sera reacted with pig gastric vesicles enriched in H+/K(+)-ATPase (gastric proton pump). Immunoblotting experiments indicated that the alpha subunit of the H+/K(+)-ATPase was the major antigen in the vesicles with two of the sera reacting strongly. We further characterized the specificity of the antibodies using partial sequences of the pig alpha subunit fused with truncated TrpE (anthranilate synthase). The antibodies from autoimmune gastritis patients reacted differently to the two fusion proteins (Met-1 to Ala-79, and Arg-606 to Ile-964), indicating that each patient sera contains a mixture of autoantibodies recognizing different epitopes with variable contents.
采用免疫学方法分析了来自自身免疫性胃炎患者、含有抗胃壁细胞自身抗体的人血清。酶联免疫吸附测定表明,所有9份血清均与富含H⁺/K⁺-ATP酶(胃质子泵)的猪胃小泡发生反应。免疫印迹实验表明,H⁺/K⁺-ATP酶的α亚基是小泡中的主要抗原,其中两份血清与之强烈反应。我们使用与截短的色氨酸合成酶E(邻氨基苯甲酸合酶)融合的猪α亚基的部分序列,进一步鉴定了抗体的特异性。来自自身免疫性胃炎患者的抗体对两种融合蛋白(Met-1至Ala-79,以及Arg-606至Ile-964)的反应不同,这表明每位患者的血清都含有识别不同表位、含量各异的自身抗体混合物。