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Comparison of suspension MDCK cells, adherent MDCK cells, and LLC-MK2 cells for selective isolation of influenza viruses to be used as vaccine seeds

tetano

Editor, Senior Moderator
Influenza Other Respir Viruses. 2019 Oct 25. doi: 10.1111/irv.12694. [Epub ahead of print] [h=1]Comparison of suspension MDCK cells, adherent MDCK cells, and LLC-MK2 cells for selective isolation of influenza viruses to be used as vaccine seeds.[/h]
Harada Y[SUP]1[/SUP], Takahashi H[SUP]1[/SUP], Trusheim H[SUP]2[/SUP], Roth B[SUP]2[/SUP], Mizuta K[SUP]3[/SUP], Hirata-Saito A[SUP]4[/SUP], Ogane T[SUP]4[/SUP], Odagiri T[SUP]1[/SUP], Tashiro M[SUP]1[/SUP], Yamamoto N[SUP]1,[/SUP][SUP]5[/SUP].
[h=3]Author information[/h] 1 Influenza Virus Research Center, National Institute of Infectious Diseases, Tokyo, Japan. 2 Novartis Vaccines and Diagnostics GmbH, Marburg, Germany. 3 Yamagata Prefectural Institute of Public Health, Yamagata, Japan. 4 Tochigi Prefectural Institute of Public Health and Environmental Science, Utsunomiya, Japan. 5 Department of Infection Control Science, Graduate School of Medicine, Juntendo University, Tokyo, Japan.

[h=3]Abstract[/h] [h=4]BACKGROUND:[/h] Cell-based influenza vaccines can solve the problem of the frequent occurrence of egg adaptation-associated antigenic changes observed in egg-based vaccines. Seed viruses for cell-based vaccines can be prepared from clinical specimens by cell culture; however, clinical samples risk harboring respiratory viruses other than influenza virus. Therefore, it is necessary to investigate the patterns of co-infection in clinical samples and explore whether cell culture technology can selectively propagate influenza viruses from samples containing other respiratory viruses.
[h=4]METHODS:[/h] A total of 341 clinical specimens were collected from patients with influenza or influenza-like illness and analyzed by ResPlex II assay to detect 18 respiratory viruses. The patterns of co-infection were statistically analyzed with Fisher's exact test. The samples with double or triple infections were passaged in suspension MDCK cells (MDCK-S), adherent MDCK cells (MDCK-A), and LLC-MK2D cells. Cell-passaged samples were analyzed by ResPlex II assay again to investigate whether each cell line could amplify influenza viruses and eliminate other respiratory viruses.
[h=4]RESULTS:[/h] Double infections were detected in 8.5% and triple infections in 0.9% of the collected clinical specimens. We identified four pairs of viruses with significant correlation. For all samples with double and triple infection, MDCK-S and MDCK-A could selectively propagate influenza viruses, while eliminating all contaminating viruses. In contrast, LLC-MK2D showed lower isolation efficiency for influenza virus and higher isolation efficiency for coxsackievirus/echovirus than MDCK-S and MDCK-A.
[h=4]CONCLUSIONS:[/h] Both MDCK-S and MDCK-A are considered suitable for the preparation of influenza vaccine seed viruses without adventitious agents or egg-adaptation mutations.
? 2019 The Authors. Influenza and Other Respiratory Viruses Published by John Wiley & Sons Ltd.


[h=4]KEYWORDS:[/h] Madin-Darby canine kidney cell line; adventitious virus; cell-based vaccine; influenza; vaccine seed virus

PMID: 31651085 DOI: 10.1111/irv.12694
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