Moore K M, Jackwood M W
Department of Avian Medicine, College of Veterinary Medicine, University of Georgia, Athens 30602-4875.
Avian Dis. 1994 Apr-Jun;38(2):218-24.
Production of monoclonal antibodies (MAbs) to Bordetella avium surface proteins led to further characterization of the hemagglutinin. Proteins obtained by homogenization of whole-cell B. avium were used to immunize mice for the production of monoclonal antibodies. Immunoprecipitation and Western blot techniques were used to determine the specificity of three MAbs, which all recognized the B. avium 41-kilodalton (kd) surface protein. In addition, all of the MAbs inhibited hemagglutination (HA) of guinea pig erythrocytes by B. avium. The 41-kd protein recognized by the MAbs was observed by the indirect immunofluorescence test and sodium dodecyl sulfate-polyacrylamide gel electrophoresis to bind to guinea pig erythrocytes. When HA-positive isolates of B. avium were treated with periodic acid, which cleaves carbohydrates from proteins, the isolates became HA-negative. Likewise, treatment of HA-positive B. avium isolates with proteinase K, which would also remove carbohydrates associated with proteins on the surface of the bacterium, inhibited HA. Considering these data, we suggest that the B. avium hemagglutinin is a carbohydrate closely associated with the 41-kd surface protein.
针对鸟博德特氏菌表面蛋白生产单克隆抗体（MAb），促使对血凝素进行了进一步的特性分析。通过将鸟博德特氏菌全细胞匀浆获得的蛋白质用于免疫小鼠以生产单克隆抗体。采用免疫沉淀和蛋白质印迹技术来确定三种单克隆抗体的特异性，这三种单克隆抗体均识别鸟博德特氏菌41千道尔顿（kd）的表面蛋白。此外，所有单克隆抗体均抑制鸟博德特氏菌对豚鼠红细胞的血凝作用（HA）。通过间接免疫荧光试验和十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳观察到，单克隆抗体识别的41-kd蛋白与豚鼠红细胞结合。当用高碘酸（可从蛋白质上裂解碳水化合物）处理血凝阳性的鸟博德特氏菌分离株时，这些分离株变为血凝阴性。同样，用蛋白酶K处理血凝阳性的鸟博德特氏菌分离株（蛋白酶K也会去除与细菌表面蛋白质相关的碳水化合物）也会抑制血凝。基于这些数据，我们认为鸟博德特氏菌血凝素是一种与41-kd表面蛋白紧密相关的碳水化合物。
