Characterization of high-growth reassortant influenza A viruses generated in MDCK cells cultured in serum-free medium.
In the present study reassortant influenza A viruses of both the H1N1 and H3N2 type were generated in Madin Darby Canine Kidney cells grown in the absence of fetal bovine serum (MDCK-SF1 cells). To this end, MDCK-SF1 cells were simultaneously infected with one of the high-growth laboratory strains A/Puerto Rico/8/34 (H1N1) or A/Hong Kong/2/68 (H3N2) and recent H3N2 and H1N1 vaccine strains, respectively. Reassortant viruses obtained from these mixed infections were genetically characterized by RT-PCR and restriction enzyme analysis and their growth properties were compared to those of the corresponding field strains. Reassortant H3N2 viruses inherited the matrix and polymerase pa gene whilst H1N1 reassortant viruses inherited the matrix and polymerase pbl gene of the high-growth parent. Reassortant viruses generally gave higher viral yields, as measured by a haemagglutination assay, than their wild type counterparts. The procedure followed results in the generation of high-growth reassortant viruses in weeks. The use of MDCK-SF1 cells together with these reassortants for generating influenza virus antigens can significantly speed up the vaccine production procedure.
Voeten
J T
JT
Institute of Virology and WHO National Influenza Centre, Erasmus University Rotterdam, The Netherlands. voeten@viro.fgg.eur.nl
Brands
R
R
Palache
A M
AM
van Scharrenburg
G J
GJ
Rimmelzwaan
G F
GF
Osterhaus
A D
AD
Claas
E C
EC
D000818:Animals; D002460:Cell Line; D016895:Culture Media, Serum-Free; D004285:Dogs; D006385:Hemagglutination Inhibition Tests; D006386:Hemagglutination Tests; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000378:metabolism; D009980:Influenza A virus / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D009439:Neuraminidase / Q000235:genetics / Q000378:metabolism; D016865:Reassortant Viruses / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D015183:Restriction Mapping; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014763:Viral Matrix Proteins / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication
D016895:Culture Media, Serum-Free; D006389:Hemagglutinins, Viral; D008562:Membrane Glycoproteins; D014763:Viral Matrix Proteins; D014764:Viral Proteins; C077346:influenza virus polymerase basic protein 1; D009439:Neuraminidase