West Nile virus (WNV) is a globally disseminated Flavivirus that is associated with encephalitis outbreaks in humans and horses. The continuous global outbreaks of West Nile disease in the bird, human, and horse populations, with no preventative measures for humans, pose a major public health threat. The development of a vaccine that contributes to the “One Health” Initiative could be the answer to prevent the spread of the virus and control human and animal disease. The current commercially available veterinary vaccines are generally costly and most require high levels of biosafety for their manufacture. Consequently, we explored making a particulate vaccine candidate made transiently in plants as a more cost-effective and safer means of production. A WNV virus-like particle-display-based vaccine candidate was generated by the use of the SpyTag/SpyCatcher (ST/SC) conjugation system. The WNV envelope protein domain III (EDIII), which contains WNV-specific epitopes, was fused to and displayed on AP205 phage virus-like particles (VLPs) following the production of both separately in Nicotiana benthamiana. Co-purification of AP205 and EDIII genetically fused to ST and SC, respectively, resulted in the conjugated VLPs displaying EDIII with an average coupling efficiency of 51%. Subcutaneous immunisation of mice with 5 μg of purified AP205: EDIII VLPs elicited a potent IgG response to WNV EDIII. This study presents the potential plants being used as biofactories for making significant pharmaceutical products for the “One Health” Initiative and could be used to address the need for their local production in low- and middle-income countries (LMICs).

西尼罗河病毒 (WNV) 是一种全球传播的病毒 黄病毒这与人类和马的脑炎爆发有关。西尼罗河病在鸟类、人类和马群中的全球持续爆发，对人类没有任何预防措施，构成了重大的公共卫生威胁。开发有助于“同一个健康”倡议的疫苗可能是防止病毒传播和控制人类和动物疾病的答案。目前市售的兽用疫苗通常价格昂贵，并且大多数需要高水平的生物安全性来生产。因此，我们探索了一种在植物中瞬时制备的颗粒状候选疫苗，作为一种更具成本效益和更安全的生产方式。使用 SpyTag/SpyCatcher (ST/SC) 结合系统生成了基于 WNV 病毒样颗粒展示的候选疫苗。本氏烟草. 分别与 ST 和 SC 遗传融合的 AP205 和 EDIII 的共纯化导致缀合的 VLP 显示 EDIII，平均偶联效率为 51%。用 5 μg 纯化的 AP205: EDIII VLP 对小鼠进行皮下免疫，引发了对 WNV EDIII 的强效 IgG 反应。本研究介绍了用作生物工厂的潜在植物，用于为“同一个健康”计划制造重要的医药产品，并可用于解决低收入和中等收入国家 (LMIC) 本地生产的需求。