Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Generation of a High-Growth Influenza Vaccine Strain in MDCK Cells for Vaccine Preparedness

  • Kim, Eun-Ha (Microbiology Department, College of Medicine and Medical Research Institute, Chungbuk National University) ;
  • Kwon, Hyeok-Il (Microbiology Department, College of Medicine and Medical Research Institute, Chungbuk National University) ;
  • Park, Su-Jin (Microbiology Department, College of Medicine and Medical Research Institute, Chungbuk National University) ;
  • Kim, Young-Il (Microbiology Department, College of Medicine and Medical Research Institute, Chungbuk National University) ;
  • Si, Young-Jae (Microbiology Department, College of Medicine and Medical Research Institute, Chungbuk National University) ;
  • Lee, In-Won (Microbiology Department, College of Medicine and Medical Research Institute, Chungbuk National University) ;
  • Kim, Se mi (Microbiology Department, College of Medicine and Medical Research Institute, Chungbuk National University) ;
  • Kim, Soo-In (Virus Vaccine, Green Cross Research Center) ;
  • Ahn, Dong-Ho (Virus Vaccine, Green Cross Research Center) ;
  • Choi, Young-Ki (Microbiology Department, College of Medicine and Medical Research Institute, Chungbuk National University)
  • Received : 2017.12.05
  • Accepted : 2018.03.31
  • Published : 2018.06.28
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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 demands 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 $10^8PFU/ml$. Genetic characterization revealed that these viruses mainly had amino acid substitutions in internal genes and exhibited higher polymerase activities. By using a series of Rg viruses, we demonstrated the essential residues of each gene and identified a set of high-growth strains in MDCK cells ($PB1_{D153N}$, $M1_{A137T}$, and $NS1_{N176S}$). 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 $10^7PFU/ml$) in both attached- and suspended-MDCK cells compared with 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 that are cheap and can be grown promptly and vigorously as a substitute for 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.

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