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Neutralizing Monoclonal Antibody to Edema Toxin and Its Effect on Murine Anthrax

机译：中和水肿毒素单克隆抗体及其对小鼠炭疽的作用

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Edema factor (EF) is a component of an anthrax toxin that functions as an adenylate cyclase. Numerous monoclonal antibodies (MAbs) have been reported for the other Bacillus anthracis toxin components, but relatively few to EF have been studied. We report the generation of six murine hybridoma lines producing two IgM and four IgG1 MAbs to EF. Of the six MAbs, only one IgM neutralized EF, as assayed by an increase in cyclic AMP (cAMP) production by Chinese hamster ovary (CHO) cells. Analysis of the variable gene elements revealed that the single neutralizing MAb had a different binding site than the others. There was no competition between the neutralizing IgM and the nonneutralizing IgG MAbs indicative of different specificity. MAb-based capture enzyme-linked immunosorbent assay (ELISA) detected EF in liver lysates from mice infected with B. anthracis Sterne 34F2. Administration of the neutralizing IgM MAb to A/JCr mice lethally infected with B. anthracis strain Sterne had no significant effect on median time to death, but mice treated with the MAb were more likely to survive infection. Combining the neutralizing IgM to EF with a subprotective dose of a neutralizing MAb to protective antigen (PA) prolonged mean time to death of infected mice, suggesting that neutralization of EF and PA could produce synergistic beneficial effects. In summary, the results from our study and literature observations suggest that the majority of Abs to EF are nonneutralizing, but the toxin has some epitopes that can be targeted by the humoral response to generate useful Abs that may contribute to defense against anthrax.
机译：水肿因子（EF）是炭疽毒素的一种成分，起着腺苷酸环化酶的作用。其他炭疽芽孢杆菌毒素成分的单克隆抗体（MAb）已有报道，但针对EF的研究相对较少。我们报告了产生两个IgM和四个IgG1 MAb到EF的六个小鼠杂交瘤细胞系的生成。通过中国仓鼠卵巢（CHO）细胞循环AMP（cAMP）产量的增加，在六种单克隆抗体中，只有一种IgM中和了EF。对可变基因元件的分析显示，单个中和性单克隆抗体的结合位点与其他结合位点不同。中和性IgM和非中和性IgG MAb之间没有竞争，表明存在不同的特异性。基于MAb的捕获酶联免疫吸附测定（ELISA）在感染 B的小鼠的肝裂解物中检测到EF。炭疽菌Sterne 34F2。将中和的IgM MAb给予致命感染了 B的A / JCr小鼠。炭疽热斯特恩菌株对中位死亡时间没有显着影响，但接受单抗的小鼠更有可能幸免于感染。将中和的IgM与EF结合使用亚保护剂量的中和性单克隆抗体与保护性抗原（PA），可延长感染小鼠的平均死亡时间，这表明EF和PA的中和作用可产生协同有益作用。总之，我们的研究结果和文献观察表明，大多数针对EF的Abs均未中和，但该毒素具有一些表位，可以通过体液反应靶向该表位，从而产生有用的Abs，从而有助于抵抗炭疽病。

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