Vaccine efficacy induced by 2020-2021 seasonal influenza-derived H3N1 virus-like particles co-expressing M2e5x or N2.

Influenza A matrix protein 2 (M2e) and neuraminidase (NA) antigens are known to play important roles in mounting a broad range of protection. Nonetheless, the protective efficacy of the VLP vaccines co-expressing both M2e and NA antigens has not been explored. In this study, we generated 2020/2021 seasonal influenza H3N1 VLPs that co-expressed either M2e5x (H3N1M2e5x) or N2 (H3N1N2 VLP) antigens. The protective efficacy of these VLPs was assessed by challenge infection with heterologous H3N2 and heterosubtypic H1N1 and H5N1 viruses in mice. Both VLP formulations induced cross-protection against distinct viruses, H3N1M2e5x VLPs elicited higher levels of cross-reactive IgG in sera against H1N1 and H5N1 viruses than H3N1N2 VLPs. Compared to H3N1N2 VLPs, H3N1M2e5x VLPs also induced substantially enhanced germinal center B cell responses while inhibiting IFN-γ production in the lungs. Importantly, H3N1M2e5x VLPs significantly reduced the lung virus titers upon H1N1, H3N2, and H5N1 challenge infections. These results indicated that VLPs comprising the M2e5x antigen are a promising vaccine design strategy that could aid in the pursuit of a universal influenza vaccine.