Suitability of PER.C6 cells to generate epidemic and pandemic influenza vaccine strains by reverse genetics.
Reverse genetics, the generation of influenza viruses from cDNA, presents a rapid method for creating vaccine strains. The technique necessitates the use of cultured cells. Due to technical and regulatory requirements, the choice of cell lines for production of human influenza vaccines is limited. PER.C6 cells, among the most extensively characterized and documented cells, support growth of all influenza viruses tested to date, and can be grown to high densities in large bioreactors in the absence of serum or micro carriers. Here, the suitability of these cells for the generation of influenza viruses by reverse genetics was investigated. A range of viruses reflective of vaccine strains was rescued exclusively using PER.C6 cells by various transfection methods, including an animal component-free procedure. Furthermore, a whole inactivated vaccine carrying the HA and NA segments of A/HK/156/97 (H5N1) that was both rescued from and propagated on PER.C6 cells, conferred protection in a mouse model. Thus PER.C6 cells provide an attractive platform for generation of influenza vaccine strains via reverse genetics.
Koudstaal
W
W
Crucell Holland BV, Leiden, The Netherlands. wouter.koudstaal@crucell.com
Hartgroves
L
L
Havenga
M
M
Legastelois
I
I
Ophorst
C
C
Sieuwerts
M
M
Zuijdgeest
D
D
Vogels
R
R
Custers
J
J
de Boer-Luijtze
E
E
de Leeuw
O
O
Cornelissen
L
L
Goudsmit
J
J
Barclay
W
W
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D018929:Cell Culture Techniques; D002460:Cell Line; D005260:Female; D006385:Hemagglutination Inhibition Tests; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000254:growth & development* / Q000276:immunology; D053122:Influenza A Virus, H3N2 Subtype / Q000235:genetics / Q000254:growth & development* / Q000276:immunology; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000254:growth & development* / Q000276:immunology; D007252:Influenza Vaccines / Q000235:genetics* / Q000276:immunology; D007251:Influenza, Human / Q000517:prevention & control; D051379:Mice; D008807:Mice, Inbred BALB C; D016865:Reassortant Viruses / Q000235:genetics / Q000254:growth & development* / Q000276:immunology; D012720:Severity of Illness Index; D016019:Survival Analysis; D014162:Transfection / Q000379:methods; D010948:Viral Plaque Assay
D000914:Antibodies, Viral; D007252:Influenza Vaccines