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Establishment of MDCK Stable Cell Lines Expressing TMPRSS2 and MSPL and Their Applications in Propagating Influenza Vaccine Viruses in Absence of Exog

tetano

Editor, Senior Moderator
Biotechnol Res Int. 2015;2015:402628. doi: 10.1155/2015/402628. Epub 2015 Mar 30.
[h=1]Establishment of MDCK Stable Cell Lines Expressing TMPRSS2 and MSPL and Their Applications in Propagating Influenza Vaccine Viruses in Absence of Exogenous Trypsin.[/h] Wen Z[SUP]1[/SUP], Wu C[SUP]1[/SUP], Chen W[SUP]1[/SUP], Zeng X[SUP]1[/SUP], Shi J[SUP]1[/SUP], Ge J[SUP]1[/SUP], Chen H[SUP]1[/SUP], Bu Z[SUP]1[/SUP].
[h=3]Author information[/h]

[h=3]Abstract[/h] We established two Madin-Darby canine kidney (MDCK) cell lines stably expressing human airway transmembrane protease: transmembrane protease, serine 2 (TMPRSS2) and mosaic serine protease large form (MSPL) which support multicycle growth of two H5 highly pathogenic avian influenza viruses (HPAIV) recombinant vaccines (Re-5 and Re-6) and an H9 avian influenza virus (AIV) recombinant vaccine (Re-9) in the absence of trypsin. Data showed that the cell lines stably expressed TMPRSS2 and MSPL after 20 serial passages. Both MDCK-TMPRSS2 and MDCK-MSPL could proteolytically cleave the HA of Re-5, Re-6, and Re-9 and supported high-titer growth of the vaccine without exogenous trypsin. Re-5, Re-6, and Re-9 efficiently infected and replicated within MDCK-TMPRSS2 and MDCK-MSPL cells and viral titer were comparable to the virus grown in MDCK cells with TPCK-trypsin. Thus, our results indicate a potential application for these cell lines in cell-based influenza vaccine production and may serve as a useful tool for HA proteolytic cleavage-related studies.


PMID: 25918647 [PubMed]
 
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