Vibrio cholerae causes cholera and other infections, especially in children under five years of age. Cholera toxin (CT), toxin-coregulated pilus (TCP) and outer membrane protein W (OmpW) are three major virulence factors of this bacterium. The emergence of antimicrobial-resistant (AMR) strains and the absence of a comprehensive and flawless vaccine, has prompted other treatments. There are several advantages of egg yolk antibodies (IgY) over other immunotherapy agents, such as economic feasibility, high yield simple production, and better immune responsiveness to mammalian antigens due to phylogenetic distance. Accordingly, in the present study, IgYs against recombinant proteins CtxB (responsible for the CT binding to eukaryotic cells), TcpA (enhances bacterial attachment to enterocytes) and OmpW were produced, in single, coupled or combined forms, to evaluate and compare their protectivity potency. Immunoreactivity of IgYs were examined through protein and whole cell ELISA, their specificity was confirmed by western blotting, and their neutralizing effects on CT was evaluated in Y1 cell culture. Produced IgYs were gavage administered to different groups of infant mice infected with V. cholerae. The results indicated that IgYs produced against CtxB had the highest titers, and were able to neutralize cytotoxicity effects in Y1 cell culture, while the highest protection in the mice challenge was obtained by IgY-TcpA. No considerable increase was observed in immunoreactivity or protectivity of antibodies produced against combined antigens. The produced IgYs showed a good antigen-specificity and protectivity which can be used in passive immunotherapy against cholera.

霍乱弧菌引起霍乱和其他感染，尤其是在五岁以下的儿童中。霍乱毒素（CT），毒素核心菌毛（TCP）和外膜蛋白W（OmpW）是该细菌的三个主要毒力因子。抗菌素耐药性（AMR）菌株的出现以及缺乏全面，无缺陷的疫苗的出现，促使其他治疗方法的出现。卵黄抗体（IgY）相对于其他免疫治疗剂有多个优势，例如经济可行性，高产量的简单生产以及由于系统发生距离而对哺乳动物抗原具有更好的免疫反应能力。因此，在本研究中，针对重组蛋白CtxB（负责CT与真核细胞的结合），TcpA（增强细菌与肠上皮细胞的附着）和OmpW的IgY以单，偶联或组合形式生产，评估和比较其保护效力。通过蛋白质和全细胞ELISA检查IgY的免疫反应性，通过蛋白质印迹证实其特异性，并在Y1细胞培养物中评估其对CT的中和作用。将产生的IgY灌胃给不同组的被感染的婴儿小鼠霍乱弧菌。结果表明，针对CtxB产生的IgYs具有最高的效价，并且能够中和Y1细胞培养物中的细胞毒性作用，而通过IgY-TcpA获得的小鼠攻击中的最高保护作用。没有观察到针对结合抗原产生的抗体的免疫反应性或保护性的显着提高。产生的IgYs显示出良好的抗原特异性和保护性，可用于针对霍乱的被动免疫疗法。