Adenoviral-vectored universal influenza vaccines administered intranasally reduce lung inflammatory responses upon viral challenge 15 months post-vaccination

ABSTRACT New approaches to vaccination are needed for protection against rapidly evolving viruses because emerging variant strains often evade neutralizing antibodies. Alternative strategies include targeting highly conserved antigens that change more slowly over time, which could provide the basis for universal vaccines. Recombinant adenovirus (rAd)-based vaccines expressing conserved influenza virus antigens nucleoprotein (NP) and matrix 2 (M2) provide powerful, long-lasting protection against challenge with diverse influenza strains, especially if administered intranasally. After intranasal administration of these rAd-based vaccines, animals are protected against morbidity and mortality in part by strong local T cell responses. However, concerns have been raised that these local responses could result in immune-mediated lung damage. In this study, we vaccinated mice intranasally with rAd vectors expressing influenza A/NP and/or M2, or influenza B/NP as a control, and then challenged them with an influenza A virus 15 months later. Protection persisted, as shown by accelerated viral clearance. Vaccination reduced the incidence and/or severity of histopathologic findings associated with influenza infection. Vaccination with A/NP+M2-rAd or A/NP-rAd reduced cytokine responses in the lungs following challenge. Granzyme B levels peaked and resolved earlier in animals vaccinated with A/NP+M2-rAd or A/NP-rAd than in other groups, consistent with a rapid CD8 T cell response that resolved more quickly. An influx of neutrophils, shown by a rise in neutrophil elastase, occurred in all groups following the challenge but returned to baseline more quickly in the two A/NP-vaccinated groups. Thus, vaccines inducing potent local immunity in the respiratory tract provided protection, rather than damage, to the lungs. IMPORTANCE Vaccines targeting highly conserved proteins can protect broadly against diverse viral strains. When a vaccine is administered to the respiratory tract, protection against disease is especially powerful. However, it is important to establish that this approach is safe. When vaccinated animals later encounter viruses, does reactivation of powerful local immunity, including T cell responses, damage the lungs? This study investigates the safety of mucosal vaccination of the respiratory tract. Non-replicating adenoviral vaccine vectors expressing conserved influenza virus proteins were given intranasally. This vaccine-induced protection persists for at least 15 months. Vaccination did not exacerbate inflammatory responses or tissue damage upon influenza virus infection. Instead, vaccination with nucleoprotein reduced cytokine responses and histopathology, while neutrophil and T cell responses resolved earlier. The results are promising for safe vaccination at the site of infection and thus have implications for the control of influenza and other respiratory viruses. Vaccines targeting highly conserved proteins can protect broadly against diverse viral strains. When a vaccine is administered to the respiratory tract, protection against disease is especially powerful. However, it is important to establish that this approach is safe. When vaccinated animals later encounter viruses, does reactivation of powerful local immunity, including T cell responses, damage the lungs? This study investigates the safety of mucosal vaccination of the respiratory tract. Non-replicating adenoviral vaccine vectors expressing conserved influenza virus proteins were given intranasally. This vaccine-induced protection persists for at least 15 months. Vaccination did not exacerbate inflammatory responses or tissue damage upon influenza virus infection. Instead, vaccination with nucleoprotein reduced cytokine responses and histopathology, while neutrophil and T cell responses resolved earlier. The results are promising for safe vaccination at the site of infection and thus have implications for the control of influenza and other respiratory viruses.