Abstract
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).
Highlights
West Nile virus (WNV) is a member of the genus Flavivirus in the family Flaviviridae (Lindenbach et al, 2007)
A higher yield of soluble WNV-envelope protein domain III (EDIII)-SC was observed in leaves co-infiltrated with CRT in comparison to CNX as assessed by the intensity of the protein band obtained at 4 dpi
We evaluated several co-extraction ratios of leaves infiltrated with ST-AP205 to leaves infiltrated with WNV-EDIII-SC/CRT and found that the increase in fresh leaf weight (FLW) of the latter resulted in a decrease in uncoupled ST-AP205 coat protein (CP) subunits, suggesting an increase in the coupling efficiency (Supplementary Figure 7)
Summary
West Nile virus (WNV) is a member of the genus Flavivirus in the family Flaviviridae (Lindenbach et al, 2007). Several equine West Nile vaccines (whole inactivated virus, inactivated/live recombinant) produced in the United States (Castro-Jorge et al, 2019) and Europe (Rebollo et al, 2018) are licenced for use. These vaccines can be difficult and expensive to obtain in low- and middle-income countries (LMICs), with regards to import permits, cost of the vaccines, and the need for annual vaccination. The “One Health” Initiative is dedicated to improving the lives of all species, both human and animal, through the integration of human and veterinary medicine and environmental science
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