In the present study, we carried-out assessment of efficacy of different immunization strategies using two bivalent vaccine formulations containing antigens of inactivated Newcastle disease virus (NDV-genotype VIId) and reassortant highly pathogenic avian influenza virus (H5N1-HPAIV) mixed with Montanide ISA71 and Montanide Gel02 as adjuvants. The efficacy of the prepared vaccines was evaluated by determining the cellular and humoral immune responses. In addition, protection against H5N1-AIV and NDV-genotype VIId challenge viruses post vaccination was assessed when Montanide-Gel02 based vaccine was inoculated in 10-days-old specific pathogen free chicks intraocularly once, twice or once followed by a boost with the Montanide ISA71 based vaccine. The cytokines profile analysis demonstrated that the prime-boost strategy induced the highest up-regulation in interferon-gamma (11.39-fold change) and interleukin-6 (14.12-fold change) genes expression. Also, enhanced lymphocytes proliferation was recorded beside increased antibody titers with protection levels reaching 50 and 60% against H5N1 and NDV challenge; respectively. Immunization with Montanide ISA71 inactivated vaccine induced 80% protection; however, the prime-boost combination afforded complete protection (100%) in the challenged chickens against mortality, clinical signs and virus shedding. Finally, these results highlight the significance of considering not only different vaccine platforms but also vaccination strategies to maximize protection against AIV and NDV with regards to the longevity of the vaccine-induced immune response.
Keywords: Genotype VIId; H5N1; Montanide; Newcastle disease virus; Prime-boost.