We developed a chicken mucosa-targeting polymeric nanoparticle vaccine (~90 nm) co-delivering viral antigenCpG adjuvant using a water-in-oil-in-water double emulsion technique. The particles are biodegradablebiocompatible. This nanoparticle significantly stimulated chicken Harderian gland B cell proliferationsubsequent local IgAIgG secretion in tears. The mucosal Ig protected chickens from virus infection by lowering virus replication at the infection sitethereafter reducing tracheal epithelial ciliostasissystemic infection, relieving clinical symptoms.

We developed a polymeric nanoparticle vaccine (~90 nm) co-delivering viral antigenCpG adjuvant using a water-in-oil-in-water double emulsion technique. The particles are biodegradablebiocompatible. This is the first vaccine to evaluate  mucosal immune response of the chicken Harderian gland, including antigen-secreting cell stainingimmune-related gene expression screening. Overall, this mucosal nanoparticle vaccine shows promising immunostimulatory effects on the chicken respiratory immune system, enabling effective mucosal immunity against an avian coronavirus.

We are able to produce the protein antigen using a 10-liter bioreactor,subsequently purified by the automated purification system. The batch yield has now reached 0.2 g of recombinant RBD protein, which is sufficient for preparing 5000 vaccine doses. Furthermore, the nanoparticle vaccines can be lyophilized for long-term storage without the cold-chain issues. This vaccine shows a great potential for commercializationbeing used in the field. As avian coronavirus is an economically important respiratory pathogen worldwide, this vaccine is expected to have a global market.

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