Microbiology, Department of Biopharmaceutical Sciences, Gifu Pharmaceutical University, Gifu, Japan.
PLoS One. 2011;6(10):e26526. doi: 10.1371/journal.pone.0026526. Epub 2011 Oct 19.
Shiga toxins (Stxs) are involved in the development of severe systemic complications associated with enterohemorrhagic Escherichia coli (EHEC) infection. Various neutralizing agents against Stxs are under investigation for management of EHEC infection. In this study, we immunized chickens with formalin-inactivated Stx-1 or Stx-2, and obtained immunoglobulin Y (IgY) from the egg yolk. Anti-Stx-1 IgY and anti-Stx-2 IgY recognized the corresponding Stx A subunit and polymeric but not monomeric B subunit. Anti-Stx-1 IgY and anti-Stx-2 IgY suppressed the cytotoxicity of Stx-1 and Stx-2 to HeLa 229 cells, without cross-suppressive activity. The suppressive activity of these IgY was abrogated by pre-incubation with the corresponding recombinant B subunit, which suggests that the antibodies directed to the polymeric B subunits were predominantly involved in the suppression. In vivo, the intraperitoneal or intravenous administration of these IgY rescued mice from death caused by intraperitoneal injection of the corresponding toxin at a lethal dose. Moreover, oral administration of anti-Stx-2 IgY reduced the mortality of mice infected intestinally with EHEC O157:H7. Our results therefore suggest that anti-Stx IgY antibodies may be considered as preventive agents for Stx-mediated diseases in EHEC infection.
志贺毒素(Stx)参与了与肠出血性大肠杆菌(EHEC)感染相关的严重全身并发症的发展。目前正在研究各种针对 Stx 的中和剂来治疗 EHEC 感染。在这项研究中,我们用甲醛灭活的 Stx-1 或 Stx-2 免疫鸡,并从蛋黄中获得免疫球蛋白 Y(IgY)。抗 Stx-1 IgY 和抗 Stx-2 IgY 识别相应的 Stx A 亚单位和聚合但非单体 B 亚单位。抗 Stx-1 IgY 和抗 Stx-2 IgY 抑制了 Stx-1 和 Stx-2 对 HeLa 229 细胞的细胞毒性,没有交叉抑制活性。这些 IgY 的抑制活性被与相应重组 B 亚单位预孵育所消除,这表明针对聚合 B 亚单位的抗体主要参与了抑制作用。在体内,这些 IgY 的腹腔内或静脉内给药可挽救小鼠免于因腹腔内注射相应毒素致死剂量而死亡。此外,口服抗 Stx-2 IgY 可降低经肠道感染 EHEC O157:H7 的小鼠的死亡率。因此,我们的研究结果表明,抗 Stx IgY 抗体可被视为 EHEC 感染中 Stx 介导疾病的预防剂。