Construction with recombinant epitope-expressing baculovirus enhances protective effects of inactivated H9N2 vaccine against heterologous virus.

Although the use of inactivated vaccines has kept avian influenza (AI) outbreaks largely under control, they fail to prevent virus shedding. To enhance the efficacy of inactivated H9N2 AIV vaccines (InV), we constructed a multi-epitope recombinant baculovirus (BV-BNT) containing two B cell epitopes and nine T cell epitopes of H9N2 AIV for combined immunization with InV. The results showed that HI titer, IgG and IgM levels, and the percentage of B cells, CD4 T cells, CD8 T cells, and CD4CD8 T cells were significantly higher in the InV+BV-BNT immunization group than the InV immunization group. Besides, the expression levels of IL-1β, IFN-γ, IFN-α, IL-4, IL-13, and CXCLi1 were significantly higher in the InV+BV-BNT group than the InV group. Moreover, four conservative peptides (NP, NP, NS, and NP) significantly stimulated splenocytes to express IFN-γ in the InV+BV-BNT group instead of InV group. After heterologous virus challenging, the percentages of CD4 T and CD8 T cells were significantly upregulated in the InV+BV-BNT group compared to Inv group at 3 DPI. Viral loads in oropharyngeal of the InV+BV-BNT group was significantly lower than that in the InV group at 3 days post-infection (DPI). Furthermore, compared to the InV group, the virus positivity rate of oropharyngeal and cloacal swabs in the InV+BV-BNT group was lower at 5 DPI, with none positive at 7 DPI. Hence, this study indicated that the combined immunization of InV and BV-BNT could induce stronger humoral and cellular immune responses, shorten the detoxification period and reduce viral load compared to Inv alone, which suggests BV-BNT could act as a supplementary vaccine to potentially address the protection deficiency of the H9N2 inactivated vaccine.