Nakai T, Horiguchi Y, Kume K
Research Center for Veterinary Science, Kitasato Institute, Chiba, Japan.
Nihon Juigaku Zasshi. 1990 Jun;52(3):533-42. doi: 10.1292/jvms1939.52.533.
Four hybridoma cell lines producing monoclonal antibodies (MAbs) against Actinobacillus (Haemophilus) pleuropneumoniae were established by fusion of mouse myeloma and spleen cells obtained from mice immunized with a serotype 2, strain SH-15. Enzyme-linked immunosorbent assay-inhibition tests with antigens obtained from 12 serotype strains of A. pleuropneumoniae and 9 other gram-negative bacteria showed that all the MAbs bound to only serotype 2 strains of A. pleuropneumoniae. The epitopes recognized by the MAbs were located on a carbohydrate moiety of lipopolysaccharide (LPS) of the organism, which was sensitive to periodate oxidation. In immunoblotting analyses of LPS obtained from A. pleuropneumoniae serotype 2, all the four MAbs reacted specifically with the characteristic ladder bands of LPS detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These results suggest that O-antigen side chains of the LPS are one of the antigenic determinants responsible for the serotype-specificity of A. pleuropneumoniae.
通过将小鼠骨髓瘤细胞与用2型SH - 15菌株免疫的小鼠脾脏细胞融合，建立了4株产生抗胸膜肺炎放线杆菌（嗜血杆菌）单克隆抗体（MAb）的杂交瘤细胞系。用从12株胸膜肺炎放线杆菌血清型菌株和9种其他革兰氏阴性菌获得的抗原进行酶联免疫吸附抑制试验，结果表明所有单克隆抗体仅与胸膜肺炎放线杆菌2型菌株结合。单克隆抗体识别的表位位于该菌脂多糖（LPS）的碳水化合物部分，对高碘酸盐氧化敏感。在对胸膜肺炎放线杆菌2型获得的LPS进行免疫印迹分析时，所有4种单克隆抗体都与十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳检测到的LPS特征梯状条带发生特异性反应。这些结果表明，LPS的O抗原侧链是胸膜肺炎放线杆菌血清型特异性的抗原决定簇之一。
