CHARACTERIZATION AND COMPARATIVE BACTERICIDAL ACTIVITY OF MONOCLONAL-ANTIBODIES TO BORDETELLA-PERTUSSIS LIPO-OLIGOSACCHARIDE-A
CHARACTERIZATION AND COMPARATIVE BACTERICIDAL ACTIVITY OF MONOCLONAL-ANTIBODIES TO BORDETELLA-PERTUSSIS LIPO-OLIGOSACCHARIDE-A
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DOI：
10.1099/00221287-137-4-905
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发表时间：
1991-04-01
期刊：
影响因子：
--
通讯作者：
BRODEUR, BR
中科院分区：
文献类型：
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作者：
ARCHAMBAULT, D;RONDEAU, P;BRODEUR, BR
用百日咳杆菌n -月桂醇肌氨酸膜提取物（SME）免疫小鼠脾细胞，制备产生单克隆抗体（mab）的杂交瘤细胞株。通过SDS-PAGE对SME或纯化的LOS进行免疫印迹，发现7个单抗对百日咳脂寡糖（LOS）具有特异性。所有单克隆抗体均与LOS AB表型的百日咳Tohama I株发生反应，而与LOS B表型的非典型变异株134无反应。经蛋白酶K处理后，单克隆抗体的免疫反应性保持不变，而经高碘酸钠处理后，单克隆抗体的免疫反应性丧失。在对副百日咳杆菌和其他革兰氏阴性菌进行检测时，未观察到与单克隆抗体的交叉反应性。然而，所有单克隆抗体都能与支杆菌反应。与活百日咳细胞的结合试验表明，单克隆抗体与细胞表面暴露的抗原决定因子强烈反应。其中5个单抗具有较高的细菌细胞裂解能力。用蛋白A纯化的浓度在0.22 - 2.2 μ g mAb ml-1之间（每450mu -l测定0.1 -1 μ g），可以杀死至少50%的细菌。生物素化抗体的竞争免疫分析表明，抗细菌抗体和非抗细菌抗体针对百日咳B. loa的不同抗原表位。
Spleen cells from mice immunized with a Bordetella pertussis N-lauroyl sarcosine membrane extract (SME) were used to generate hybridoma cells lines producing monoclonal antibodies (mAbs). Seven mAbs were shown to be specific to B. pertussis lipo-oligosaccharide (LOS) by immunoblotting of SME or purified LOS following SDS-PAGE. All mAbs reacted with the B. pertussis Tohama I strain of the LOS AB phenotype, and did not react with the atypical variant strain 134 of the LOS B phenotype. The immune reactivity of the mAbs was retained after treatment of SME with proteinase K and was lost after sodium periodate treatment. No cross-reactivity was observed with the mAbs when tested against B. parapertussis and other Gram-negative bacteria. However, all mAbs reacted with B. bronchiseptica. Binding assays with live B. pertussis cells demonstrated that mAbs strongly reacted with cell surface exposed antigenic determinants. High bacterial cell lytic capability was observed for five of these mAbs. Concentrations between 0.22 and 2.2-mu-g mAb ml-1 (0.1 and 1-mu-g per 450-mu-l assay) purified by protein A were required to kill at least 50% of the bacteria. Competition immunoassays with biotinylated antibodies showed that the bacteriolytic and non-bacteriolytic mAbs were directed to different epitopes of the B. pertussis LOS A.