Multivalent display of anthrax toxin inhibitors

炭疽芽孢杆菌会产生一种毒素，会导致感染者的细胞死亡和死亡。即使细菌感染本身是由抗生素控制的，感染者通常也会死亡，因为炭疽杆菌体内产生的炭疽毒素(AT)与人类细胞结合，被内吞，并杀死细胞。因此，开发能阻止AT与人体细胞结合的特异性抗毒素对于有效治疗炭疽暴露是必要的。这就是本计划项目拨款的目标。J.Young和J.Collier的研究小组已经发现并表征了AT与其细胞受体之间的相互作用。这项工作推动了基于多肽的新型炭疽抗毒素的发现和开发(项目1和2)。在这里，我们建议以植物病毒颗粒豌豆花叶病毒(CPMV)或昆虫病毒鸡舍病毒(FHV)为展示平台(项目3-5)，以多价形式展示抑制肽和基于AT受体的可溶性蛋白。这些多价平台将被生产出来，并在体外和体内测试它们作为抗毒素的有效性。这些病毒耐热耐酸，容易大量生产。这两种病毒在基因、生物化学和结构上都有很好的特征，可以通过工程或化学修饰在其表面显示多个外来免疫原性或抗毒素多肽序列。曼彻斯特研究小组(项目3)此前已经证明，通过在CPMV上展示抑制肽，在体外和体内都可以有效地阻止病毒与细胞受体的相互作用。使用这一令人兴奋的结果作为概念的证明，我们将测试多价的假设 在CPMV和FHV上展示AT抑制肽将导致一系列新的有效的炭疽抗毒素和疫苗试剂的产生。

Bacillus anthracis produces a toxin that causes cell death and mortality in infected individuals. Even if the bacterial infection itself is controlled by antibiotics, an infected person often dies because the anthrax toxin (AT) produced in vivo by B. anthracis binds to human cells, is endocytosed, and kills the cells. Therefore development of specific antitoxins that prevent AT from binding to human cells is necessary for effectively treating anthrax exposure,. This is the goal of this Program Project grant. The groups of J. Young and J. Collier have discovered and characterized the interaction between AT and its cellular receptors. This work fuels the discovery and development of novel peptide-based anthrax anti-toxins (Projects 1 and 2). Here we propose to display the inhibitory peptides and soluble AT receptor-based proteins in a multivalent form using the plant virus particle cowpea mosaic virus (CPMV) or the insect virus Flock house virus (FHV) as display platforms (Projects 3-5). These multivalent platforms will be produced and tested for their efficacy as antitoxins both in vitro and in vivo. These viruses are heat and acid-stable and are easy to produce in large quantities. Both viruses have been very well characterized genetically, biochemically and structurally and can either be engineered, or chemically modified, to display multiple copies of foreign immunogenic or antitoxin peptide sequences on their surface. The Manchester group (project 3) has previously shown that a virus-cellular receptor interaction can be efficiently blocked, both in vitro and in vivo, by the multivalent display of an inhibitory peptide on CPMV. Using this exciting result as a proof of concept, we will test the hypothesis that multivalent display of AT inhibitory peptides on CPMV and FHV will lead to the creation of a panel of new and effective anthrax antitoxins and vaccine reagents.