Production and characterization of antibodies against each of the three subunits of the Bacillus cereus nonhemolytic enterotoxin complex
Production and characterization of antibodies against each of the three subunits of the Bacillus cereus nonhemolytic enterotoxin complex
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DOI：
10.1128/aem.71.12.8214-8220.2005
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发表时间：
2005-12-01
影响因子：
4.4
通讯作者：
Märtlbauer, E
中科院分区：
文献类型：
--
作者：
Dietrich, R;Moravek, M;Märtlbauer, E
非溶血性肠毒素（nonhemolytic enterotoxin，Nhe）是引起蜡样芽孢杆菌食物中毒综合征的两种三组分肠毒素之一。为了便于检测这种毒素，由亚基NheA，NheB和NheC组成，建立并表征了针对三种组分中每一种的一整套高亲和力抗体。使用偶联至用于免疫的蛋白质载体的相应合成肽产生对NheC的C末端部分（15个氨基酸）特异的兔抗血清。使用纯化的B。以蜡状芽孢杆菌胞外蛋白为免疫原，获得了一株抗NheA的单克隆抗体和几株抗NheB的单克隆抗体。与B不同菌株产生的其他蛋白无交叉反应性。观察到蜡状体。针对NheB组分的抗体能够中和Nhe的细胞毒性活性（高达98%）。基于间接酶免疫分析，本研究中开发的抗体被成功地用于表征几种B的肠毒性活性。蜡状芽孢杆菌菌株。这是第一次可以证明携带nhe基因的菌株通常表达包括NheC在内的三种组分的完整集合。然而，不同菌株产生的毒素量差异很大。
The nonhemolytic enterotoxin (Nhe) is one of the two three-component enterotoxins which are responsible for diarrheal food poisoning syndrome caused by Bacillus cereus. To facilitate the detection of this toxin, consisting of the subunits NheA, NheB, and NheC, a complete set of high-affinity antibodies against each of the three components was established and characterized. A rabbit antiserum specific for the C-terminal part (15 amino acids) of NheC was produced using a respective synthetic peptide coupled to a protein carrier for immunization. Using purified B. cereus exoprotein preparations as immunogens, one monoclonal antibody against NheA and several antibodies against NheB were obtained. No cross-reactivity with other proteins produced by different strains of B. cereus was observed. Antibodies against the NheB component were able to neutralize the cytotoxic activity (up to 98%) of Nhe. Based on indirect enzyme immunoassays, the antibodies developed in this study were successfully used in the characterization of the enterotoxic activity of several B. cereus strains. For the first time, it could be shown that strains carrying the nhe genes usually express the complete set of the three components, including NheC. However, the amount of toxin produced varies considerably between the different strains.