African green monkey kidney (Vero) cells provide an alternative host cell system for influenza A and B viruses

Abstract

The preparation of live, attenuated human influenza virus vaccines and of large quantities of inactivated vaccines after the emergence or reemergence of a pandemic influenza virus will require an alternative host cell system, because embryonated chicken eggs will likely be insufficient and suboptimal. Preliminary studies indicated that an African green monkey kidney cell line (Vero) is a suitable system for the primary isolation and cultivation of influenza A viruses (E. A. Govorkova, N. V. Kaverin, L. V. Gubareva, B. Meignier, and R. G. Webster, J. Infect. Dis. 172:250-253, 1995). We now demonstrate for the first time that Vero cells are suitable for isolation and productive replication of influenza B viruses and determine the biological and genetic properties of both influenza A and B viruses in Vero cells; additionally, we characterize the receptors on Vero cells compared with those on Madin-Darby canine kidney (MDCK) cells. Sequence analysis indicated that the hemagglutinin of Vero cell-derived influenza B viruses was identical to that of MDCK-grown counterparts but differed from that of egg-grown viruses at amino acid positions 196 to 198. Fluorescence-activated cell sorting analysis showed that although Vero cells possess predominantly ␣2,3 galactose-linked sialic acid, they are fully susceptible to infection with either human influenza A or B viruses. Moreover, all virus-specific polypeptides were synthesized in the same proportions in Vero cells as in MDCK cells. Electron microscopic and immunofluorescence studies confirmed that infected Vero cells undergo the same morphological changes as do other polarized epithelial cells. Taken together, these results indicate that Vero cell lines could serve as an alternative host system for the cultivation of influenza A and B viruses, providing adequate quantities of either virus to meet the vaccine requirements imposed by an emerging pandemic.

MDCK and Vero cells for influenza virus vaccine production: a one-to-one comparison up to lab-scale bioreactor cultivation

“…In SC medium, however, all trypsin concentrations tested resulted in similar HA profiles. Multiple additions of trypsin as described by Govorkova et al (1995Govorkova et al ( , 1996, did not result in improvement of HA titers. As this method would complicate largescale production processes, a single addition of trypsin at time of infection should be preferred.…”

“…For the development of production processes using serum-free media, these inhibitors could have a negative effect on virus production when washing and medium exchange steps are omitted before infection. While virus replication in MDCK cells seems to be only marginally effected by protease inhibitors, multiple trypsin additions have been described for Vero cells to overcome trypsin inactivation (Govorkova et al 1996).…”

“…However, influenza virus replication has not been found to be as sensitive to trypsin concentration as in Vero cells, and trypsin is usually added at time of infection only (Tree et al 2001;Genzel et al 2004Genzel et al , 2006aSchwartz et al 2006). Finally, virus adaptation to the producer cell line has often been described for the adaptation from eggs to cells (Lin et al 1997;Genzel and Reichl 2007;Govorkova et al 1996). Up to 20 passages were needed to obtain acceptable HA virus titers.…”

Alternative Experimental Models for Studying Influenza Proteins, Host–Virus Interactions and Anti-Influenza Drugs

“…These cells are widely used for protein expression studies due to the ease of transfection into these cells [84]. Vero cells are kidney cells derived from Cercopithecus aethiops (African green monkey) kidney and have been extensively used in virology research [85,86].…”

Tools to study pathogen-host interactions in bats

“…Virus isolation is generally attempted in established immortalized cell lines, such as the type I interferon deficient BHK-21 (baby Syrian hamster kidney cells) (Otsuki et al, 1979) or Vero E6 cell lines (African green monkey kidney cells) (Govorkova et al, 1996). Different primary cells from bats have been studied and some immortalized cell lines have also been generated (Table 1).…”