Abstract
Influenza virus is a negative segmented RNA virus without DNA intermediate. This makes it safer as a vaccine delivery vector than most DNA viruses that have potential to integrate their genetic elements into host genomes. In this study, we developed a universal influenza viral vector, expressing the receptor binding subdomain of botulinum neurotoxin A (BoNT/A). We tested the growth characters of the engineered influenza virus in chicken eggs and Madin–Darby canine kidney epithelial cells (MDCK), and showed that it can be produced to a titer of 5 × 106 plaque forming unites/ml in chicken eggs and MDCK cells. Subsequently, mice intranasally vaccinated with the engineered influenza virus conferred protection against challenge with lethal doses of active BoNT/A toxin and influenza virus. Our results demonstrated the feasibility to develop a dual purpose nasal vaccine against both botulism and influenza.
Highlights
Influenza A virus is a member of family orthomyxoviridae, which contains a segmented RNA genome
GENERATION OF ENGINEERED INFLUENZA VIRUSES First, we synthesized a gene segment coding 2A peptide followed by a multiple cloning site [19]
To demonstrate the use of this universal influenza viral vector, a 618 bp gene segment of botulinum neurotoxin A (BoNT/A) HC50-rbsd was inserted into engineered NA gene segment
Summary
Influenza A virus is a member of family orthomyxoviridae, which contains a segmented RNA genome. The most important application of this system is creation of live-attenuated vaccine or the generation of influenza vector to express foreign antigens [1,2,3]. In early stage of influenza virus vector development, influenza virus non-structural gene (NS) segment was engineered to express foreign antigen with deletion of the nuclear export protein (NEP), formally referred to as NS2 [7,8,9,10,11]. With the finding of 2A cleavage sequence of picornavirus, NS gene segment was modified with multi-cistronic sites. It enlarged the containment of foreign gene, kept a high growth of engineered influenza virus, and enhanced genetic stability [7, 12]
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