Cell Line Development by Systems Biology
Within an interdisciplinary project which combines the expertise of two industrial and five academic partners it is our aim to provide an innovative and highly productive cell culture-based influenza vaccine manufacturing platform. Therefore, we want to optimize influenza virus replication in well-defined human designer cells supplied by ProBioGen AG (Berlin) by manipulating the expression level of cellular factors involved in virus-host cell interactions. Based on results obtained by a genome-wide RNAi screen , different cell lines with specific genetic manipulations are being produced at the Helmholtz Centre for Infection Research (HZI, Braunschweig) and at the Max Planck Institute for Infection Biology (MPIIB, Berlin). All modified cell lines that show an improved virus production will be analyzed in more detail to understand underlying mechanisms. In addition, our cooperation partners at the Otto von Guericke University (OvGU, Magdeburg) are developing mathematical models on the single cell and on the cell population scale to quantitatively assess the impact of cell line modifications on virus yields in bioreactors. After validation and refinement of these models by experimental approaches, they shall enable us to develop strategies for the rational design of high-yield vaccine manufacturing processes.
Aim of the project
The main focus of this subproject is the thorough characterization of influenza virus propagation in genetically manipulated cell lines. Therefore, we will use a broad panel of recent influenza vaccine strains and state-of-the-art analytical tools, i.e. real-time RT-qPCR and imaging flow cytometry, to study how genetic modifications affect virus replication. Moreover, we analyze the impact of the innate immune response of human cells on viral titers, and we optimize process parameters such as cell density at time of infection, multiplicity of infection, and harvest time to improve the process yields. Combining these findings we hope to establish a novel high-productive influenza vaccine manufacturing platform based on genetically engineered human cells.
 Karlas, A., Machuy, N., Shin, Y., Pleissner, K. P., Artarini, A., Heuer, D., Becker, D., Khalil, H., Ogilvie, L. A., Hess, S., Maurer, A. P., Muller, E., Wolff, T., Rudel, T., Meyer, T. F. (2010). "Genome-wide RNAi screen identifies human host factors crucial for influenza virus replication." Nature 463(7282): 818-822.
 Watanabe, T., Watanabe, S., Kawaoka, Y. (2010). "Cellular networks involved in the influenza virus life cycle." Cell Host Microbe 7(6): 427-439.