University of Calgary, Calgary, Alberta, Canada.
Zyme Fast Inc., Oak Bank, Manitoba, Canada.
J Med Microbiol. 2011 Aug;60(Pt 8):1181-1187. doi: 10.1099/jmm.0.029835-0. Epub 2011 Apr 7.
Herein we present evidence for the therapeutic potential of colonization factor (CF)-specific egg yolk antibodies (IgY) for potentially treating acute and recurring Clostridium difficile infection (CDI) in humans. The study involved cloning, expressing as 6×His-tagged proteins in Escherichia coli, and Ni-affinity purifying three previously identified CFs (FliC, FliD and Cwp84) from C. difficile. The recombinant CF antigens were then used to immunize Leghorn chickens and CF-specific IgY antibodies were prepared from their eggs. The specificity and titre of the resulting C. difficile CF-specific IgY antibodies were assessed by ELISA and Western immunoblotting techniques. The antibodies were also screened for their ability to inhibit C. difficile adherence to human colon-derived T84 cells, and, based on these findings, one of them (FliD-specific IgY) was evaluated for its potential to prevent C. difficile-mediated morbidity and mortality in Syrian hamsters. The results revealed that purified FliD-specific IgY significantly protected hamsters from C. difficile strain 630 infection relative to control animals treated with carbonate buffer alone or IgY produced from unimmunized chicken eggs. The results suggest that egg yolk preparations obtained from chickens immunized with recombinant C. difficile CFs may represent another safe and cost-effective treatment option in humans suffering from acute or recurring CDI.
在此，我们提出了定植因子（CF）特异性卵黄抗体（IgY）治疗人类急性和复发性艰难梭菌感染（CDI）的潜力的证据。该研究涉及克隆、在大肠杆菌中表达为 6×His 标记蛋白，并从艰难梭菌中镍亲和纯化三种先前鉴定的 CF（FliC、FliD 和 Cwp84）。然后，使用重组 CF 抗原免疫来亨鸡，并从其鸡蛋中制备 CF 特异性 IgY 抗体。通过 ELISA 和 Western 免疫印迹技术评估所得艰难梭菌 CF 特异性 IgY 抗体的特异性和效价。还筛选了这些抗体抑制艰难梭菌与人结肠衍生的 T84 细胞粘附的能力，并且基于这些发现，评估了其中一种（FliD 特异性 IgY）预防叙利亚仓鼠中艰难梭菌介导的发病率和死亡率的潜力。结果表明，与单独用碳酸盐缓冲液或未免疫鸡蛋产生的 IgY 处理的对照动物相比，纯化的 FliD 特异性 IgY 显著保护了仓鼠免受艰难梭菌 630 株感染。结果表明，用重组艰难梭菌 CF 免疫的鸡获得的卵黄制剂可能代表了另一种安全且具有成本效益的人类急性或复发性 CDI 治疗选择。
