Aim
The FLUINNATE objectives focus on the identification of influenza A virus genes and gene products which contribute to virulence/pathogenicity in experimental animal and tissue culture models. The required animals are available and will comprise mice with the wild-type Mx1 gene as part of the full innate immune response (
Picture 1), various strains with targeted mutations in specific genes and pigs as natural hosts and “mixing vessels” for influenza A viruses. In addition, human airway and porcine epithelial cell cultures will be established and characterized. Human airway epithelial cell cultures are a rare but most precious substrate to study the biology of influenza virus infection ( Picture 2). The influenza viruses used will be human, avian and swine strains, some of which will be generated by reverse genetics entirely from plasmids. Stock viruses and single and multi-segment reassortants will be produced and fully characterized together with the parental strains with respect to growth kinetics in tissue culture and in vivo, the capacity to induce or respond to interferon, and the capacity to induce disease or death in experimental animals. The technology for expressing, purifying and analyzing the viral RNA polymerase complex will be established and further refined, as well as biochemical and biophysical approaches to identify co-purifying host cellular factors. Advanced tests for protein-protein interactions such as the yeast three-hybrid system will be set up and candidate interactors evaluated in functional tests, based on transfection experiments.
 |
PROBLEM |  |
POTENTIAL APPLICATIONS |