Dupuy P, LePendu J, Jothy S, Wilkinson R D
Department of Dermatology, Royal Victoria Hospital, McGill University, Montreal, Quebec, Canada.
Hybridoma. 1990 Dec;9(6):589-96. doi: 10.1089/hyb.1990.9.589.
We report the preparation and characterization of an IgG2 monoclonal antibody (MAb), HSMA, prepared against a human pooled sweat extract (HPSE). The major component of HPSE was a mucin-type molecule, as revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) stained with periodic acid-Schiff (PAS) reagent. By immunoblotting, HSMA revealed a smear in the high molecular weight range, typical of mucins. In enzyme-linked immunosorbent assay (ELISA), HSMA failed to react with HPSE fractions isolated after anionic exchange gel chromatography. Similarly, radio-immunobinding assays demonstrated no reactivity between HSMA and A, B, H, and Lewis blood group-related structures. The immunohistological labeling on normal skin showed that HSMA reacted with the cells of eccrine sweat glands, and to a lesser extent, with sebaceous glands and epidermal cells. Periodate treatment in situ abolished these reactions, thus suggesting the carbohydrate structure of the HSMA-epitope. In indirect immunofluorescence (IF) studies, HSMA also reacted with other exocrine glands, e.g. mammary glands, sublingual glands, mixed sero-mucous glands of the trachea, and in the pancreas. Sparse positive cells were also observed in the testis, kidney, thyroid and digestive tract.
我们报告了一种IgG2单克隆抗体（MAb）HSMA的制备及特性，该抗体是针对人混合汗液提取物（HPSE）制备的。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）经高碘酸-希夫（PAS）试剂染色显示，HPSE的主要成分是一种粘蛋白型分子。通过免疫印迹法，HSMA在高分子量范围内呈现出条带涂抹现象，这是粘蛋白的典型特征。在酶联免疫吸附测定（ELISA）中，HSMA未能与阴离子交换凝胶色谱分离后的HPSE组分发生反应。同样，放射免疫结合测定表明HSMA与A、B、H以及Lewis血型相关结构之间无反应性。对正常皮肤进行免疫组织学标记显示，HSMA与小汗腺细胞发生反应，在较小程度上也与皮脂腺和表皮细胞发生反应。原位高碘酸盐处理消除了这些反应，因此提示了HSMA表位的碳水化合物结构。在间接免疫荧光（IF）研究中，HSMA也与其他外分泌腺发生反应，例如乳腺、舌下腺、气管的混合浆液-粘液腺以及胰腺。在睾丸、肾脏、甲状腺和消化道中也观察到了稀疏的阳性细胞。
