Dietrich R, Fella C, Strich S, Märtlbauer E
Institute for Hygiene and Technology of Food of Animal Origin, Veterinary Faculty, University of Munich, 80539 Munich, Germany.
Appl Environ Microbiol. 1999 Oct;65(10):4470-4. doi: 10.1128/AEM.65.10.4470-4474.1999.
A total of five hybridoma cell lines that produced monoclonal antibodies against the components of the hemolysin BL (HBL) enterotoxin complex and sphingomyelinase produced by Bacillus cereus were established and characterized. Monoclonal antibody 2A3 was specific for the B component, antibodies 1A12 and 8B12 were specific for the L(2) component, and antibody 1C2 was specific for the L(1) protein of the HBL enterotoxin complex. No cross-reactivity with other proteins produced by different strains of B. cereus was observed for monoclonal antibodies 2A3, 1A12, and 8B12, whereas antibody 1C2 cross-reacted with an uncharacterized protein of approximately 93 kDa and with a 39-kDa protein, which possibly represents one component of the nonhemolytic enterotoxin complex. Antibody 2A12 finally showed a distinct reactivity with B. cereus sphingomyelinase. The monoclonal antibodies developed in this study were also successfully applied in indirect enzyme immunoassays for the characterization of the enterotoxic activity of B. cereus strains. About 50% of the strains tested were capable of producing the HBL enterotoxin complex, and it could be demonstrated that all strains producing HBL were also highly cytotoxic.
共建立并鉴定了五种杂交瘤细胞系，它们产生针对蜡样芽孢杆菌产生的溶血素BL（HBL）肠毒素复合物和鞘磷脂酶成分的单克隆抗体。单克隆抗体2A3对B组分具有特异性，抗体1A12和8B12对L(2)组分具有特异性，抗体1C2对HBL肠毒素复合物的L(1)蛋白具有特异性。未观察到单克隆抗体2A3、1A12和8B12与不同蜡样芽孢杆菌菌株产生的其他蛋白质发生交叉反应，而抗体1C2与一种约93 kDa的未鉴定蛋白质和一种39 kDa的蛋白质发生交叉反应，后者可能代表非溶血肠毒素复合物的一个组分。抗体2A12最终显示出与蜡样芽孢杆菌鞘磷脂酶有明显的反应性。本研究中开发的单克隆抗体也成功应用于间接酶免疫测定，以鉴定蜡样芽孢杆菌菌株的肠毒素活性。约50%的测试菌株能够产生HBL肠毒素复合物，并且可以证明所有产生HBL的菌株也具有高度细胞毒性。
