tetano
Editor, Senior Moderator
J Microbiol Biotechnol. 2018 Apr 12. doi: 10.4014/jmb.1712.12007. [Epub ahead of print]
[h=1]Generation of a High-Growth Influenza Vaccine Strain in MDCK Cells for Vaccine Preparedness.[/h] Kim EH[SUP]1[/SUP], Kwon HI[SUP]1[/SUP], Park SJ[SUP]1[/SUP], Kim YI[SUP]1[/SUP], Si YJ[SUP]1[/SUP], Lee IW[SUP]1[/SUP], Kim SM[SUP]1[/SUP], Kim SI[SUP]2[/SUP], Ahn DH[SUP]2[/SUP], Choi YK[SUP]1[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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 10[SUP]8[/SUP] 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 10[SUP]7[/SUP]PFU/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.
[h=4]KEYWORDS:[/h] Bioreactors; Cell-derived vaccine; Influenza viruses; Madin-Darby canine kidney (MDCK)
PMID: 29642288 DOI: 10.4014/jmb.1712.12007
Free full text
[h=1]Generation of a High-Growth Influenza Vaccine Strain in MDCK Cells for Vaccine Preparedness.[/h] Kim EH[SUP]1[/SUP], Kwon HI[SUP]1[/SUP], Park SJ[SUP]1[/SUP], Kim YI[SUP]1[/SUP], Si YJ[SUP]1[/SUP], Lee IW[SUP]1[/SUP], Kim SM[SUP]1[/SUP], Kim SI[SUP]2[/SUP], Ahn DH[SUP]2[/SUP], Choi YK[SUP]1[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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 10[SUP]8[/SUP] 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 10[SUP]7[/SUP]PFU/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.
[h=4]KEYWORDS:[/h] Bioreactors; Cell-derived vaccine; Influenza viruses; Madin-Darby canine kidney (MDCK)
PMID: 29642288 DOI: 10.4014/jmb.1712.12007
Free full text