title: Metabolic effects of influenza virus infection in cultured animal cells: Intra- and extracellular metabolite profiling.
abstract: 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.
authors.affiliation: Delft University of Technology, Bioprocess Technology, Julianalaan 67, 2628 BC Delft, the Netherlands.
authors.forename: Joachim B
authors.initials: JB
authors.lastname: Ritter
authors.forename: Aljoscha S
authors.initials: AS
authors.lastname: Wahl
authors.forename: Susann
authors.initials: S
authors.lastname: Freund
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Genzel
authors.forename: Udo
authors.initials: U
authors.lastname: Reichl
mesh_terms: 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
chemical_list: D009711:Nucleotides; D019344:Lactic Acid; D005947:Glucose
references.citation: Vaccine. 2006 Aug 28;24(35-36):6074-87
references.pmid: 16781022
references.citation: Trends Biochem Sci. 2005 Mar;30(3):142-50
references.pmid: 15752986
references.citation: Mol Cell Biochem. 2004 Nov;266(1-2):191-8
references.pmid: 15646042
references.citation: J Gen Virol. 1984 Jul;65 ( Pt 7):1229-32
references.pmid: 6086816
references.citation: Proc Natl Acad Sci U S A. 1974 Oct;71(10):4129-32
references.pmid: 4372608
references.citation: Biotechnol Bioeng. 1997 Jul 20;55(2):305-16
references.pmid: 18636489
references.citation: Nature. 1956 Dec 1;178(4544):1230-1
references.pmid: 13387671
references.citation: Anal Biochem. 2004 Dec 1;335(1):119-25
references.pmid: 15519579
references.citation: Biologicals. 2008 May;36(3):145-61
references.pmid: 18561375
references.citation: Metab Eng. 2007 Jul;9(4):317-26
references.pmid: 17611135
references.citation: Biotechnol Bioeng. 2004 Oct 5;88(1):1-14
references.pmid: 15384040
references.citation: Drugs. 2008;68(11):1483-91
references.pmid: 18627206
references.citation: Arch Gesamte Virusforsch. 1972;39(1):26-34
references.pmid: 4673823
references.citation: Virology. 1959 Oct;9:244-59
references.pmid: 14441181
references.citation: Virology. 1956 Oct;2(5):637-55
references.pmid: 13371724
references.citation: Biotechnol Prog. 2008 Mar-Apr;24(2):311-20
references.pmid: 18215054
references.citation: Int Rev Immunol. 2003 Sep-Dec;22(5-6):425-49
references.pmid: 12959753
references.citation: Virology. 1961 Jan;13:68-77
references.pmid: 13756751
references.citation: Biotechnol Bioeng. 2005 Apr 5;90(1):46-58
references.pmid: 15736163
references.citation: Biotechnol Bioeng. 2006 Jul 5;94(4):667-79
references.pmid: 16570315
references.citation: Vaccine. 2006 Apr 12;24(16):3261-72
references.pmid: 16472544
references.citation: Virology. 1958 Apr;5(2):206-19
references.pmid: 13544101
references.citation: PLoS Pathog. 2006 Dec;2(12):e132
references.pmid: 17173481
references.citation: Biotechnol Prog. 2005 Jan-Feb;21(1):58-69
references.pmid: 15903241
references.citation: Biochim Biophys Acta. 2007 Oct;1772(10):1158-66
references.pmid: 17964123
references.citation: J Infect Dis. 1957 Mar-Apr;100(2):109-18
references.pmid: 13416643
references.citation: Nat Biotechnol. 2008 Oct;26(10):1179-86
references.pmid: 18820684
references.citation: Biotechnol Prog. 2008 Jan-Feb;24(1):110-9
references.pmid: 18171074
references.citation: Appl Microbiol Biotechnol. 1995 Dec;44(1-2):27-36
references.pmid: 8579834
references.citation: Int J Cancer. 1968 Mar 15;3(2):273-82
references.pmid: 4296943
references.citation: Curr Med Chem. 2003 Aug;10(16):1535-51
references.pmid: 12871125
references.citation: J Chromatogr B Analyt Technol Biomed Life Sci. 2006 Nov 7;843(2):216-26
references.pmid: 16798123
references.citation: Biochem J. 1967 May;103(2):514-27
references.pmid: 4291787
references.citation: Vaccine. 2003 May 1;21(16):1789-95
references.pmid: 12686096
references.citation: FEBS J. 2007 Mar;274(6):1393-418
references.pmid: 17302740
references.citation: Vaccine. 2004 Jun 2;22(17-18):2202-8
references.pmid: 15149778
references.citation: Virology. 1970 Aug;41(4):647-52
references.pmid: 4319783
references.citation: Anal Biochem. 2008 Feb 15;373(2):349-69
references.pmid: 18036549
references.citation: Biotechnol Bioeng. 2008 Sep 1;101(1):135-52
references.pmid: 18646224
references.citation: Arch Gesamte Virusforsch. 1967;21(3):425-36
references.pmid: 5600965
references.citation: Appl Environ Microbiol. 2008 Sep;74(18):5710-23
references.pmid: 18641162
references.citation: FEBS Lett. 1975 Feb 15;50(3):334-8
references.pmid: 1116604
references.citation: Intervirology. 1977;8(2):100-9
references.pmid: 558175
references.citation: Cell Microbiol. 2006 Mar;8(3):375-86
references.pmid: 16469051
references.citation: J Virol. 2003 Feb;77(3):2174-81
references.pmid: 12525652
references.citation: Infect Immun. 1974 May;9(5):824-7
references.pmid: 4363231
references.citation: Biochem J. 1980 Dec 15;192(3):793-800
references.pmid: 7236238
references.citation: Gene Ther. 2008 Apr;15(7):545-52
references.pmid: 18273055
references.citation: Vaccine. 2009 May 5;27(20):2712-22
references.pmid: 19428884