Establishment of MDCK Stable Cell Lines Expressing TMPRSS2 and MSPL and Their Applications in Propagating Influenza Vaccine Viruses in Absence of Exogenous Trypsin. (30th March 2015)

• Record Type: Journal Article
• Authors: Wen, Zhiyuan; Wu, Chao; Chen, Weiye; Zeng, Xianying; Shi, Jianzhong; Ge, Jinying; Chen, Hualan; Bu, Zhigao
• Other Names: Canovas Manuel Academic Editor.
• Is Part Of: Biotechnology research international. Volume 2015(2015)
• Publication Date: 2015-03-30
• Subjects: Biotechnology -- Periodicals; Biotechnology; Biotechnology; Periodicals; 660.6
• DOI: 10.1155/2015/402628
• Languages: English
• ISSNs: 2090-3138

Abstract

Abstract : 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.