Imaging of Influenza Virus Replication to Optimize Cell Culture-based Vaccine Production
Every year, vaccines against the seasonal strains of Influenza A and B viruses are produced in large quantities. These vaccines are prepared from influenza viruses that were propagated in suitable substrates such as fertilized chicken eggs or cultured mammalian cells. To maximize the yield of cell culture-based processes, high-yield producer cell lines have to be selected, and efficient virus propagation strategies have to be developed. For sustained advances in cell culture-based influenza vaccine production, it is crucial to understand and to control virus-host cell interactions that determine the amount of viruses produced.
Aim of the project
In this project, we want to gain quantitative and qualitative insights into influenza virus replication in animal cells that will help to improve industrial influenza vaccine production (Figure 1). For this, we try to identify cellular bottlenecks and to evaluate virus-host cell interactions that may limit virus replication and vaccine yields.
To identify putative limiting processes and factors in the influenza virus life cycle, we perform state-of-the-art imaging flow cytometry with fluorescent probes (e.g., antibodies, molecular beacons) to monitor proteins, nucleic acids, and other relevant cellular and viral features (e.g., glycans, protein phosphorylation states) in infected cells. We then statistically correlate the spatial-temporal expression and distribution patterns of the cellular and viral components with the productivity of the cells, what helps us to understand the basics of influenza virus vaccine production