Britta Peschel

Infection dynamics and virus-induced apoptosis in influenza virus A infected adherent and suspension MDCK cells

ISBN:

978-3-8440-3738-8

Series:

Forschungsberichte aus dem Max-Planck-Institut für Dynamik komplexer technischer Systeme
Herausgeber: Prof. Dr. Peter Benner, Prof. Dr.-Ing. Udo Reichl, Prof. Dr.-Ing. Andreas Seidel-Morgenstern and Prof. Dr.-Ing. Kai Sundmacher
Magdeburg

Volume:

Keywords:

influenza vaccine; cell culture; adherent cells; apoptosis; suspension cells; infection dynamics

Type of publication:

Thesis

Language:

English

Pages:

196 pages

Figures:

61 figures

Weight:

291 g

Format:

21 x 14,8 cm

Bindung:

Paperback

Price:

48,80 € / 61,00 SFr

Published:

July 2015

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DOI:

10.2370/9783844037388 (Online document)

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Abstract

Besides the classical production in embryonated chicken eggs, influenza vaccines can also be produced in cell culture-based processes. In order to contribute to the optimisation of these processes, aim of this thesis was to characterise influenza virus infection dynamics and virus-induced apoptosis of host cells. Flow cytometric staining of the influenza virus NP protein was used to monitor the time course of infection and a TUNEL assay to characterise progress of apoptosis.

Firstly, infections of adherent Madin-Darby canine kidney (MDCK) cells with different influenza virus strains at several multiplicity of infection (MOI) were performed, followed by studies on defective interfering particles (DIPs). Secondly, infections of adherent MDCK cells were additionally performed in stirred tank bioreactors. Finally, the MDCK.SUS2 suspension cell line was characterised for influenza virus propagation.

In summary, this work demonstrated low MOI infection conditions to be beneficial for optimisation of infectious virus yields in influenza A virus production. Infection dynamics depended, however, on virus strain, especially in terms of virus-induced apoptosis. In addition, presence of DIPs in seed virus strongly influenced infectious virus yields. Finally, a MDCK suspension cell line was demonstrated to be an alternative to adherent MDCK cells, offering a production process that is easier to scale-up. Thereby, this work contributed to the understanding of influenza virus propagation in cell culture, and provided valuable insights for optimisation of upstream processing in influenza vaccine production.