Metabolic effects of influenza virus infection in cultured animal cells: Intra- and extracellular metabolite profiling.
BACKGROUND
Many details in cell culture-derived influenza vaccine production are still poorly understood and approaches for process optimization mainly remain empirical. More insights on mammalian cell metabolism after a viral infection could give hints on limitations and cell-specific virus production capacities. A detailed metabolic characterization of an influenza infected adherent cell line (MDCK) was carried out based on extracellular and intracellular measurements of metabolite concentrations.


RESULTS
For most metabolites the comparison of infected (human influenza A/PR/8/34) and mock-infected cells showed a very similar behavior during the first 10-12 h post infection (pi). Significant changes were observed after about 12 h pi: (1) uptake of extracellular glucose and lactate release into the cell culture supernatant were clearly increased in infected cells compared to mock-infected cells. At the same time (12 h pi) intracellular metabolite concentrations of the upper part of glycolysis were significantly increased. On the contrary, nucleoside triphosphate concentrations of infected cells dropped clearly after 12 h pi. This behaviour was observed for two different human influenza A/PR/8/34 strains at slightly different time points.


CONCLUSIONS
Comparing these results with literature values for the time course of infection with same influenza strains, underline the hypothesis that influenza infection only represents a minor additional burden for host cell metabolism. The metabolic changes observed after 12 h pi are most probably caused by the onset of apoptosis in infected cells. The comparison of experimental data from two variants of the A/PR/8/34 virus strain (RKI versus NIBSC) with different productivities and infection dynamics showed comparable metabolic patterns but a clearly different timely behavior. Thus, infection dynamics are obviously reflected in host cell metabolism.
Ritter
Joachim B
JB
Delft University of Technology, Bioprocess Technology, Julianalaan 67, 2628 BC Delft, the Netherlands.
Wahl
Aljoscha S
AS
Freund
Susann
S
Genzel
Yvonne
Y
Reichl
Udo
U
D000818:Animals; D002452:Cell Count; D002478:Cells, Cultured / Q000378:metabolism* / Q000821:virology*; D005947:Glucose / Q000378:metabolism; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000378:metabolism*; D019344:Lactic Acid / Q000378:metabolism; D055432:Metabolomics / Q000379:methods*; D009711:Nucleotides / Q000378:metabolism; D010427:Pentose Phosphate Pathway / Q000502:physiology
D009711:Nucleotides; D019344:Lactic Acid; D005947:Glucose
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