Researcher

Britta Peschel
Phone:+49 391 6110 211

Sandtorstrasse 1, 39106 Magdeburg, Germany

Additional Information

Collaborations:
PSPD Group (Prof. A. Kienle);
PSE Group (Prof. K. Sundmacher, Dr. A. Voigt)

Start:
2008/10/01

End: Sept. 2013

Flow Cytometric Analysis of Virus-induced Apoptosis and Virus Replication

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Flow Cytometric Analysis of Virus-induced Apoptosis and Virus Replication

Motivation

To improve the yields of cell culture-based influenza vaccine production it is important to understand infection dynamics on a cellular level. As it is a powerful tool to investigate cell populations on a single-cell level, we are using flow cytometry as method of choice.

Because programmed cell death (apoptosis) is known to influence production yields it is a focus of our research. Using the apoptosis-detecting TUNEL assay and virus protein specific antibodies we are able to monitor four different cell populations during the infection in the bioreactor. Namely, we are able to differentiate between uninfected cells, apoptotic cells, infected cells, and cells that are infected and apoptotic at the same time [1,2]. Thus, flow cytometry allows us to characterize infection and apoptosis dynamics of different influenza vaccine strains, offering a profound insight into progress of infection at a cellular level.

Dot plot of a two color staining (apoptosis and infection status) of influenza infected MDCK cells. Ordinate: TUNEL fluorescence, abscissa: virus protein fluorescence Zoom Image
Dot plot of a two color staining (apoptosis and infection status) of influenza infected MDCK cells. Ordinate: TUNEL fluorescence, abscissa: virus protein fluorescence [less]

Aim of the project

A profound understanding of cellular processes taking place during an influenza infection is the basis for optimizing the vaccine production process. We seek to use the knowledge about apoptosis induction and virus spread for optimizing virus yields and thereby enhancing the efficiency of the process. Furthermore, these data are used to improve mathematical infection models, which allow a theoretical analysis of the process.

References

[1] Schulze-Horsel, J., Genzel, Y., Reichl, U. (2008) Flow cytometric monitoring of influenza A virus infection in MDCK cells during vaccine production. BMC Biotechnology 8:45.
[2] Schulze-Horsel, J., Schulze, M., Agalaridis, G., Genzel, Y., Reichl, U. (2009) Infection dynamics and virus-induced apoptosis in cell culture-based influenza vaccine production - Flow cytometry and mathematical modeling. Vaccine 27(20), 2712-2722.
[3] Schulze-Horsel, J. (2011) Zellphysiologische Charakterisierung von Zellkulturen in der Influenza-Impfstoffproduktion, PhD Thesis (German).

Related projects

Model-based Investigation of Influenza Virus Replication in Mammalian Cell Culture
Influenza Vaccine Production in Microcarrier Systems and Suspension Cells
Process Synthesis and Process Dynamics (Prof. Dr.-Ing. Achim Kienle)
Process Systems Engineering (Prof. Dr.-Ing. Kai Sundmacher)

 
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