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Generation of a High-Growth Influenza Vaccine Strain in MDCK Cells for Vaccine Preparedness

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  • Generation of a High-Growth Influenza Vaccine Strain in MDCK Cells for Vaccine Preparedness

    J Microbiol Biotechnol. 2018 Apr 12. doi: 10.4014/jmb.1712.12007. [Epub ahead of print]
    Generation of a High-Growth Influenza Vaccine Strain in MDCK Cells for Vaccine Preparedness.

    Kim EH1, Kwon HI1, Park SJ1, Kim YI1, Si YJ1, Lee IW1, Kim SM1, Kim SI2, Ahn DH2, Choi YK1.
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    Abstract

    As shown during the 2009 pandemic H1N1 [A(H1N1)pdm09] outbreak, egg-based influenza vaccine production technology is insufficient to meet global demand during an influenza pandemic. Therefore, there is a need to adapt cell culture-derived vaccine technology using suspended cell lines for more rapid and larger-scale vaccine production. In this study, we attempted to generate a high growth influenza vaccine strain in MDCK cells using an A/Puerto/8/1934 (H1N1) vaccine seed strain. Following 48 serial passages with four rounds of virus plaque purification in MDCK cells, we were able to select several MDCK-adapted plaques that could grow over 108 PFU/ml. Genetic characterization revealed that these viruses mainly had amino acid substitutions in internal genes and exhibited higher polymerase activities. By using of a series of Rg viruses, we demonstrated the essential residues of each gene and identified a set of high-growth strains in MDCK cells (PB1D153N, M1A137T and NS1N176S). In addition, we confirmed that in the context of the high growth A/PR/8/34 backbone, A/California/7/2009 (H1N1), A/Perth/16/2009 (H3N2), and A/environment/Korea/deltaW150/2006 (H5N1) also showed significantly enhanced growth properties (more than 107PFU/ml) in both attached- and suspended-MDCK cells compared to each representative virus and the original PR8 vaccine strain. Taken together, this study demonstrates the feasibility of a cell culture-derived approach to produce seed viruses for influenza vaccines which are cheap, promptly and vigorously growing which can substitute the egg-based vaccines.Thus, our results suggest that MDCK cell-based vaccine production is a feasible option for producing large-scale vaccines in case of pandemic outbreaks.


    KEYWORDS:

    Bioreactors; Cell-derived vaccine; Influenza viruses; Madin-Darby canine kidney (MDCK)

    PMID: 29642288 DOI: 10.4014/jmb.1712.12007
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