Rabbit nasal immunization against influenza by dry-powder form of chitosan nanospheres encapsulated with influenza whole virus and adjuvants

Abstract

Influenza virus is one of the main causes of respiratory diseases in human. Although different vaccines have been produced during past decades, there is still a huge demand for a safe influenza vaccine with the ability to induce mucosal immune responses and sufficient protection, especially in elderly patients. In this study, chitosan nanospheres were employed as the drug delivery system. Influenza virus, CpG oligodeoxynucleotide (CpG ODN) and Quillaja saponins (QS) were incorporated in this nanospheric system. Three doses of dry powder nanosphere vaccine were nasally administered to rabbits on days 0, 45 and 60, followed by a final booster injection on day 75. Both humoral and cellular immune responses were investigated. Hemagglutination inhibition (HI) antibody titer was elevated in all groups compared to the control group at the end of vaccination in rabbits receiving nanospheres loaded with virus and CpG, CH(WV+CpG) (P<0.001). Rabbit serum IgG raised significantly in all the vaccinated groups, with the highest responses in CH(WV+CpG) group. CH(WV+CpG) and CH(WV) induced significant sIgA titers (P<0.001). CpG adjuvant also showed a prominent role in the stimulation and secretion of of IL-2 and IFN-γ cytokines (3 and 3.5 fold increase, respectively). Finally, as CH(WV+CpG) depicted to be effective in induction of humoral and cellular immune responses after nasal administration, this nanoparticulate adjuvant could be identified as an efficient adjuvant/delivery system for mucosal immunization against influenza virus.