REACTION OF ESTABLISHED CELL LINES OF HUMAN LUNG CARCINOMA (A-549) AND ENDOTHELIUM (ECV-304) TO THE INFECTION WITH VARIOUS MULTIPLICITY
OF INFLUENZA VIRUS A
D.M. Danilenko,1v S.S. Smirnova,1 T.D. Smirnova,1,* M.M. Pisareva,1 M.A. Plotnikova,1
A.O. Durnova,2 M.Y. Eropkin 1
1 Research Institute of Influenza of Ministry of Health of Russia, St. Petersburg, 197022, and
2 Research Institute of Obsterics, Gynecology and Reproductology named after D.O. Ott, St. Petersburg, 199034;
* e-mail: cellcultures@influenza.spb.ru
The fate of cells infected with influenza virus depends not only on their biological properties and characteristics
of a virus but also on the intensity of infection, i. e. multiplicity of infection (moi). The aim of the current
study was to compare two cultured cell lines derived from human lung epithelium (A-549) and endothelium
(ECV-304) infected with various moi of influenza A virus. Both cell lines under study were shown to enhance
their proliferation and the induction of apoptosis as a result of infection with the low moi (1—10 infectious doses
— ID). Cells A-549 demonstrated stimulating effect of low moi viral infection at 1—2 days earlier comparing
to ECV-304 cells. Both cell lines responded with the enhancement of proliferation during 4 consecutive
passages on the infection with higher doses (1000 ID). It was shown that low moi had no stimulating effect on
cell A-549 in case if the virus H3N2 was cultured in chicken embryos while MDCK-grown virus did stimulate
cell proliferation. Immunocytochemistry permitted to detect changes in p53 and Cyclin A cellular localization
which proved to be moi-dependent. Under low moi (1 ID) the intra-nuclear localization of Cyclin A was prevailing
(63 %) while under high moi (100 ID) the intra-nuclear localization of p53 was observed (54 %). These findings
have confirmed that the low moi stimulates cell proliferation and high moi inhibited cellular division.
Overall, the current study shown low sensitivity of A-549 and ECV-304 cell lines to influenza A viruses which
had been confirmed by gradual decrease of viral RNA expression as well as decreased formation of influenza
virions after several cell culture passages. Thus, it can be speculated that induction of proliferation of cells in
A-549 and ECV-304 lines depends on low moi and the effect was confirmed by impaired formation of virus
particles or viral proteins. This study has revealed similarities and differences in the reaction of A-549 and
ECV-304 cell lines to various moi of influenza A virus and the obtained results can be attributed to the biological
properties of cells, viruses and infection dose.
Key words:
cell lines A-549 and ECV-304, influenza A virus, cell proliferation, apoptosis, infectious
dose, expression of viral RNA
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