Monoclonal antibodies specific for Escherichia coli J5 lipopolysaccharide: cross-reaction with other gram-negative bacterial species
Monoclonal antibodies specific for Escherichia coli J5 lipopolysaccharide: cross-reaction with other gram-negative bacterial species
复制标题
大肠杆菌 J5 脂多糖特异性单克隆抗体：与其他革兰氏阴性菌种的交叉反应
作者：
L. Mutharia;G. Crockford;W. C. Bogard;W. RobertE.;'. Hancock
本文报道了四株抗大肠杆菌J5的单克隆抗体。这些单克隆抗体中的每一种都与纯化的E. coli J5、深度粗糙突变沙门氏菌（Salmonella miniaturiRe595）、根癌农杆菌（Agrobacterium tumefaciens）和铜绿假单胞菌（Pseudomonas aeruginosa）PAO 1以及铜绿假单胞菌的纯化脂质A。酶联免疫吸附试验使用的外膜从各种革兰氏阴性菌表明，这些脂质A特异性单克隆抗体相互作用的84和97%之间的革兰氏阴性菌种测试。其中一种单克隆抗体5E4显示出与35种测试的外膜或脂多糖抗原中的34种相互作用。分离的铜绿假单胞菌外膜组分的蛋白质电泳印迹的免疫酶染色用于证明抗体5E4与来自所有17种血清型菌株和所有14种铜绿假单胞菌临床分离株的类似快速迁移条带相互作用，所述条带对应于粗糙脂多糖。同样，碘化山羊抗小鼠免疫球蛋白用于检测单克隆抗体8A1与来自肺炎克雷伯氏菌和光滑和粗糙的大肠杆菌菌株的纯化脂多糖的Western电泳印迹上的快速迁移条带的结合。大肠杆菌、鼠伤寒沙门氏菌和S.明尼苏达这些结果表明，相当大的保护单抗原位点的革兰氏阴性菌的脂质A。
Four monoclonal antibodies against Escherichia coli J5 were studied. Each of these monoclonal antibodies reacted with purified lipopolysaccharides from E. coli J5, the deep rough mutant Salmonella minnesota Re595, Agrobacterium tumefaciens, and Pseudomonas aeruginosa PAO1 as well as with the purified lipid A of P. aeruginosa. Enzyme-linked immunosorbent assays using the outer membranes from a variety of gram-negative bacteria demonstrated that these lipid A-specific monoclonal antibodies interacted with between 84 and 97% of the gram-negative bacterial species tested. One of the monoclonal antibodies, 5E4, was shown to interact with 34 of the 35 outer membrane or lipopolysaccharide antigens tested. Immunoenzymatic staining of Western electrophoretic blots of separated P. aeruginosa outer membrane components was used to demonstrate that antibody 5E4 interacted with a similar fast-migrating band, corresponding to rough lipopolysaccharide, from all 17 serotype strains and all 14 clinical isolates of P. aeruginosa. Similarly, iodinated goat anti-mouse immunoglobulin was used to detect the binding of monoclonal antibody 8A1 to a fast-migrating band on Western electrophoretic blots of purified lipopolysaccharides from Klebsiella pneumoniae and both smooth and rough strains of E. coli, Salmonella typhimurium, and S. minnesota. These results suggest considerable conservation of single antigenic sites in the lipid A of gram-negative bacteria.