Preparation and immunoprotective effects of a virus-like particle candidate vaccine of the dominant epidemic D3 genotype coxsackievirus A6 in China

Coxsackievirus A6 of the D3a genotype (CVA6 D3a) is a primary pathogen causingmainland of China's hand, foot, and mouth disease (HFMD). Viral-like particle (VLP) vaccines represent a potential candidate vaccine to prevent HFMD. This study collected Anti-CVA6 D3a VLPs serum from BALB/c female mice immunized using CVA6 D3a VLPs. The neutralizing antibody levels were compared against the representative 14-JX2018 (D3a) and N4-YN2015 (D3b) strains between the antisera of different immune pathways. The immunoprotective effect of anti-CVA6 D3a VLPs against these strains was monitored using pathological sections and immunohistochemical results of lung and skeletal muscle tissues in seven-day-old Institute of Cancer Research (ICR) mice. Immunological protection against different branches of viruses was evaluated in 7-day-old (serum passive immune protection) and 14-day-old (VLPs active immune protection) neonatal ICR mice models. Serum-neutralizing antibody levels were positively correlated with the number of immunizations and higher against 14-JX2018 than against N4-YN2015. Furthermore, these levels were significantly higher with abdominal injection than intramuscular injection. The immunized serum of 7-day-old ICR mice inoculated three times was 100 % protected against CVA6 D3a 14-JX2018 (lethal titer: 10^6.25 TCID₅₀) and CVA6 D3b N4-YN2015 (lethal titer: 10^5.25TCID₅₀) fatal attacks, respectively. For ICR mice that have completed two active immunizations for 14 days, both CVA6 D3a 14-JX2015 (challenge titer: 10^8.25 TCID₅₀) and CVA6 D3b N4-YN2015 (challenge titer: 10^7.25 TCID₅₀) were used for the challenge, and the mice were able to survive. Overall, the CVA6 D3a VLPs prepared in this study are a potential vaccine candidate for CVA6, as it has the optimal protective effect against both CVA6 D3a and D3b evolutionary branches viruses.