tetano
Editor, Senior Moderator
J Biotechnol. 2012 Oct 15. pii: S0168-1656(12)00680-3. doi: 10.1016/j.jbiotec.2012.10.005. [Epub ahead of print]
Effect of natural and chimeric hemagglutinin genes on influenza A virus replication in baby hamster kidney cells.
van Wielink R, Harmsen MM, Martens DE, de Leeuw OS, Peeters BP, Wijffels RH, Moormann RJ.
Source
Central Veterinary Institute of Wageningen UR (CVI), P.O. Box 65, 8200 AB Lelystad, the Netherlands; Wageningen University, Bioprocess Engineering, P.O. Box 8129, 6700 EV Wageningen, the Netherlands.
Abstract
Baby hamster kidney (BHK21) cells are used to produce vaccines against various viral veterinary diseases, including rabies and foot-and-mouth-disease. Although particular influenza virus strains replicate efficiently in BHK21 cells the general use of these cells for influenza vaccine production is prohibited by the poor replication of most strains, including model strain A/PR/8/34 [H1N1] (PR8). We now show that in contrast to PR8, the related strain A/WSN/33 [H1N1] (WSN) replicates efficiently in BHK21 cells. This difference is determined by the hemagglutinin (HA) protein since reciprocal reassortant viruses with swapped HAs behave similarly with respect to growth on BHK21 cells as the parental virus from which their HA gene is derived. The ability or inability of six other influenza virus strains to grow on BHK21 cells appears to be similarly dependent on the nature of the HA gene since reassortant PR8 viruses containing the HA of these strains grow to similar titres as the parental virus from which the HA gene was derived. However, the growth to low titres of a seventh influenza strain was not due to the nature of the HA gene since a reassortant PR8 virus containing this HA grew efficiently on BHK21 cells. Taken together, these results suggest that the HA gene often primarily determines influenza replication efficiency on BHK21 cells but that in some strains other genes are also involved. High virus titres could be obtained with reassortant PR8 strains that contained a chimeric HA consisting of the HA1 domain of PR8 and the HA2 domain of WSN. HA1 contains most antigenic sites and is therefore important for vaccine efficacy. This method of producing the HA1 domain as fusion to a heterologous HA2 domain could possibly also be used for the production of HA1 domains of other viruses to enable the use of BHK21 cells as a generic platform for veterinary influenza vaccine production.
Copyright ? 2012. Published by Elsevier B.V.
PMID:
23079079
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/23079079
Effect of natural and chimeric hemagglutinin genes on influenza A virus replication in baby hamster kidney cells.
van Wielink R, Harmsen MM, Martens DE, de Leeuw OS, Peeters BP, Wijffels RH, Moormann RJ.
Source
Central Veterinary Institute of Wageningen UR (CVI), P.O. Box 65, 8200 AB Lelystad, the Netherlands; Wageningen University, Bioprocess Engineering, P.O. Box 8129, 6700 EV Wageningen, the Netherlands.
Abstract
Baby hamster kidney (BHK21) cells are used to produce vaccines against various viral veterinary diseases, including rabies and foot-and-mouth-disease. Although particular influenza virus strains replicate efficiently in BHK21 cells the general use of these cells for influenza vaccine production is prohibited by the poor replication of most strains, including model strain A/PR/8/34 [H1N1] (PR8). We now show that in contrast to PR8, the related strain A/WSN/33 [H1N1] (WSN) replicates efficiently in BHK21 cells. This difference is determined by the hemagglutinin (HA) protein since reciprocal reassortant viruses with swapped HAs behave similarly with respect to growth on BHK21 cells as the parental virus from which their HA gene is derived. The ability or inability of six other influenza virus strains to grow on BHK21 cells appears to be similarly dependent on the nature of the HA gene since reassortant PR8 viruses containing the HA of these strains grow to similar titres as the parental virus from which the HA gene was derived. However, the growth to low titres of a seventh influenza strain was not due to the nature of the HA gene since a reassortant PR8 virus containing this HA grew efficiently on BHK21 cells. Taken together, these results suggest that the HA gene often primarily determines influenza replication efficiency on BHK21 cells but that in some strains other genes are also involved. High virus titres could be obtained with reassortant PR8 strains that contained a chimeric HA consisting of the HA1 domain of PR8 and the HA2 domain of WSN. HA1 contains most antigenic sites and is therefore important for vaccine efficacy. This method of producing the HA1 domain as fusion to a heterologous HA2 domain could possibly also be used for the production of HA1 domains of other viruses to enable the use of BHK21 cells as a generic platform for veterinary influenza vaccine production.
Copyright ? 2012. Published by Elsevier B.V.
PMID:
23079079
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/23079079