CeBiTec Distinguished Lecture


Friday, October 29th 2010, 17 c.t.


Lecture Hall H13


Prof. Dr.-Ing. Udo Reichl

Max-Planck-Institut für Dynamik komplexer technischer Systeme Otto-von-Guericke Universität Magdeburg


Influenza virus production in mammalian cell culture – from cellular metabolism, viral genome replication and proteomics to mathematical models

    In the next decades mammalian cell culture-derived influenza vaccines will play a crucial role not only in prevention and control of seasonal disease but also for pandemic preparedness and response planning. With the establishment of cell culture technologies to complement egg-based production systems significant efforts in design and optimization of upstream as well as downstream processing methods are required. High virus yields, which depend on host cell line, virus seed and cultivation conditions, are of particular interest. For process monitoring comprehensive analytical methods to characterize cell growth and virus replication are required. In addition, mathematical modeling is of crucial importance to obtain a quantitative understanding of the enormous complexity of virus host-cell interactions. For a thorough description of virus dynamics, various levels of interactions need to be considered. Besides virus spreading in populations of cells, virus replication within single cells has to be investigated. Furthermore, the physiological status of host cells during cell growth and, in particular, at the time of infection has to be characterized. Based on experimental data obtained from quantitative and semi-quantitative assays, hypotheses concerning time course and control of virus replication, bottlenecks for virus synthesis and cellular defense mechanisms can be formulated. Eventually, these hypotheses can be incorporated into mathematical models to support design and optimization of cell culture-derived
vaccine manufacturing. As an example for our approach, influenza A virus replication in Madin-Darby Canine Kidney (MDCK) cells is examined. The status of cells and virus replication is monitored by a number of assays to analyze the time course of extra- and intracellular metabolite concentrations (HPLC), the regulation of viral mRNA synthesis (quantitative PCR), the changes in protein expression (2D-DIGE), the degree of infection and virus-induced apoptosis (flow cytometry) and the impact of infection on various signal transduction pathways (quantitative PCR, Western Blots). Experimental data is used to establish mathematical models covering basic intracellularreplication dynamics and virus spreading in populations of cells.

Prof. Dr. Thomas Noll