Differential activation of host cell signalling pathways through infection with two variants of influenza A/Puerto Rico/8/34 (H1N1) in MDCK cells

Heynisch, B.; Frensing, Timo; Heinze, Kurt; Seitz, C.; Genzel, Yvonne; Reichl, Udo

doi:10.1016/j.vaccine.2010.07.076

Cited by 16 publications

(12 citation statements)

References 47 publications

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“…In contrast, the initial virus seed population (passage 1) of the NIBSC-strain of the adaptation series comprises more subpopulations concerning the HA-encoding RNA segment 4 than the RKI virus seed. Previously, these two (theoretical identical) influenza virus seeds have been described to differ significantly in infection characteristics such as interferon (IFN) and apoptosis induction, expression of interferon stimulated genes, final virus yields [48], [49], and the activation of general host cell response [50], [51]: PR/8/34 from NIBSC was characterized to induce higher levels of IFN and Mx expression, to induce apoptosis earlier, and to reach lower final titers than the PR/8/34 from RKI. Seitz et al hypothesized that two amino acid substitutions in the non-structural protein 1 (NS1) might be related to these differences.…”

Section: Resultsmentioning

confidence: 99%

Impact of Host Cell Line Adaptation on Quasispecies Composition and Glycosylation of Influenza A Virus Hemagglutinin

et al. 2011

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The genome of influenza A viruses is constantly changing (genetic drift) resulting in small, gradual changes in viral proteins. Alterations within antibody recognition sites of the viral membrane glycoproteins hemagglutinin (HA) and neuraminidase (NA) result in an antigenetic drift, which requires the seasonal update of human influenza virus vaccines. Generally, virus adaptation is necessary to obtain sufficiently high virus yields in cell culture-derived vaccine manufacturing. In this study detailed HA N-glycosylation pattern analysis was combined with in-depth pyrosequencing analysis of the virus genomic RNA. Forward and backward adaptation from Madin-Darby Canine Kidney (MDCK) cells to African green monkey kidney (Vero) cells was investigated for two closely related influenza A virus PR/8/34 (H1N1) strains: from the National Institute for Biological Standards and Control (NIBSC) or the Robert Koch Institute (RKI). Furthermore, stability of HA N-glycosylation patterns over ten consecutive passages and different harvest time points is demonstrated. Adaptation to Vero cells finally allowed efficient influenza A virus replication in Vero cells. In contrast, during back-adaptation the virus replicated well from the very beginning. HA N-glycosylation patterns were cell line dependent and stabilized fast within one (NIBSC-derived virus) or two (RKI-derived virus) successive passages during adaptation processes. However, during adaptation new virus variants were detected. These variants carried “rescue” mutations on the genomic level within the HA stem region, which result in amino acid substitutions. These substitutions finally allowed sufficient virus replication in the new host system. According to adaptation pressure the composition of the virus populations varied. In Vero cells a selection for “rescue” variants was characteristic. After back-adaptation to MDCK cells some variants persisted at indifferent frequencies, others slowly diminished and even dropped below the detection limit.

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Section: Resultsmentioning

confidence: 99%

Impact of Host Cell Line Adaptation on Quasispecies Composition and Glycosylation of Influenza A Virus Hemagglutinin

et al. 2011

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“…Although the mechanistic elements of influenza viral replication and production within a cell are well described, only a few studies have investigated the infection dynamic processes at the macroscopic level in large‐scale cell cultures. Recently, some process parameters were reviewed 20, 21. One important parameter for viral vaccine production is the multiplicity of infection (MOI).…”

Section: Introductionmentioning

confidence: 99%

Real‐time monitoring of influenza virus production kinetics in HEK293 cell cultures

Petiot

Kamen

2012

Biotechnology Progress

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There is an increased interest from the vaccine industry to use mammalian cell cultures for influenza vaccine manufacturing. Therefore, it became important to study the influenza infection mechanism, the viral–host interaction, and the replication kinetics from a bioprocessing stand point to maximize the influenza viral production yield in cell culture. In the present work, influenza replication kinetics was studied in HEK293 cells. Two infection conditions were evaluated, a low (0.01) and a high multiplicity of infection (1.0). Critical time points of the viral production cycle (infection, protein synthesis, viral assembly and budding, viral release, and host‐cell death) were identified in small‐scale cell cultures. Additionally, cell growth, viability, and viral titers were monitored in the viral production process. The infection state of the cultivated cell population was assessed by influenza immunolabeling throughout the culture period. Influenza virus production kinetics were also on‐line monitored by dielectric spectroscopy and successfully correlated to real‐time capacitance measures. Overall, this work provided insights into the mechanisms associated with the infection of human HEK293 cell line by the influenza virus and demonstrated, once again, the usefulness of multifrequency scanning permittivity for in‐line monitoring and supervision of cell‐based viral production processes. Published 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2013

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“…A minimal overlap between screens was observed, but core pathways were found to be conserved. The minimal overlap between different virus strains is expected since the tempo of signal transduction and host gene expression is differentially induced by different virus strains which is linked to differences in replication dynamics and virus yield [110]. It is also likely that different influenza viruses may use alternate pathways for virus replication as host pathways were generally found to be conserved between screens [72].…”

Section: Discussionmentioning

confidence: 99%

Identification of Host Kinase Genes Required for Influenza Virus Replication and the Regulatory Role of MicroRNAs

et al. 2013

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Human protein kinases (HPKs) have profound effects on cellular responses. To better understand the role of HPKs and the signaling networks that influence influenza virus replication, a small interfering RNA (siRNA) screen of 720 HPKs was performed. From the screen, 17 HPKs (NPR2, MAP3K1, DYRK3, EPHA6, TPK1, PDK2, EXOSC10, NEK8, PLK4, SGK3, NEK3, PANK4, ITPKB, CDC2L5 (CDK13), CALM2, PKN3, and HK2) were validated as essential for A/WSN/33 influenza virus replication, and 6 HPKs (CDK13, HK2, NEK8, PANK4, PLK4 and SGK3) were identified as vital for both A/WSN/33 and A/New Caledonia/20/99 influenza virus replication. These HPKs were found to affect multiple host pathways and regulated by miRNAs induced during infection. Using a panel of miRNA agonists and antagonists, miR-149* was found to regulate NEK8 expression, miR-548d-3p was found to regulate MAPK1 transcript expression, and miRs -1228 and -138 to regulate CDK13 expression. Up-regulation of miR-34c induced PLK4 transcript and protein expression and enhanced influenza virus replication, while miR-34c inhibition reduced viral replication. These findings identify HPKs important for influenza viral replication and show the miRNAs that govern their expression.

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Differential activation of host cell signalling pathways through infection with two variants of influenza A/Puerto Rico/8/34 (H1N1) in MDCK cells

Cited by 16 publications

References 47 publications

Impact of Host Cell Line Adaptation on Quasispecies Composition and Glycosylation of Influenza A Virus Hemagglutinin

Impact of Host Cell Line Adaptation on Quasispecies Composition and Glycosylation of Influenza A Virus Hemagglutinin

Real‐time monitoring of influenza virus production kinetics in HEK293 cell cultures

Identification of Host Kinase Genes Required for Influenza Virus Replication and the Regulatory Role of MicroRNAs

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