Generation of influenza A virus live vaccine

Live attenuated influenza virus vaccines (LAIV) present new possibilities for the prevention and control of influenza. Based an our expertise with the generation of recombinant baculovirus, gene expression in insect cells and the attenuation of influenza A virus, we suggest a concept for fast and easy production of influenza attenuated live vaccine. In principle, 8 influenza A virus like particles (VLPs) will be produced in high yield by the baculovirus insect cell expression system. Each of these VLPs consists of all_ Influenza A virus proteins and contains one particular influenza A virus gene (viral RNA). Thus, only when co-infected, rescue of infectious influenza virus occurs. This procedure allows fast exchange of the antigenic proteins hemagglutinin and neuraminidase, which are the basis for every influenza vaccine. The non-structural gene (NS) of influenza A virus will be truncated, thus, providing the attenuated phenotype. Co-infection will be done in Vero cells, where small amounts of VLPs (combination of 8) will yield replicating, attenuated influenza A virus, containing the desired type of hemagglutinin and neuraminidase at the surface. The baculovirus-insect cell expression system will be used to generate influenza A virus like particles. Once the required VLPs have been produced in large scale, they are available for fast and easy vaccine production. Thus, the gene transfer into mammalian cells occurs by infection, which is the most efficient way of delivery. No special expertise or complicated procedures are required for virus rescue. There is no limitation to apply this strategy to influenza A virus, bot it can be used to generate influenza B and C virus as well. Although vaccine design and production are of high interest to our research, further characterisation of basic gene properties (e.g. non-structural protein encoding gene), especially for influenza B virus, is part of our goal as well. Being able to quickly exchange and manipulate single genes of the influenza virus genome provides a versatile research tool for accelerated experiments and faster insight.