Protection against multiple influenza A virus subtypes by intranasal administration of recombinant nucleoprotein

Vaccination is a cost-effective way to control the influenza epidemic. Vaccines based on highly conserved antigens can provide protection against different influenza A strains and subtypes. In this study, the recombinant nucleoprotein (rNP) of the A/PR/8/34 (H1N1) influenza virus strain was effectively expressed using a prokaryotic expression system and then purified with a nickel-charged Sepharose affinity column as a candidate component for an influenza vaccine. The rNP was administered intranasally three times at 3-week intervals to female BALB/c mice in combination with an adjuvant (cholera toxin B subunit containing 0.2% of the whole toxin). Twenty-one days after the last immunization, the mice were challenged with homologous or heterologous influenza viruses at a lethal dose. The results showed that intranasal immunization of 10 μg rNP with adjuvant completely protected the immunized mice against the homologous influenza virus, and immunization with 100 μg rNP in combination with adjuvant provided good cross-protection against heterologous H5N1 and H9N2 avian influenza viruses. The results indicate that such a vaccine administered intranasally can induce mucosal and cell-mediated immunity, thus having the potential to control epidemics caused by new emerging influenza viruses.