Exp Mol Med.  2010 Feb;42(2):116-121. 10.3858/emm.2010.42.2.013.

Reverse genetic platform for inactivated and live-attenuated influenza vaccine

• Jung EJ
• Lee KH
• Seong BL

• KMID: 983984
• DOI: http://doi.org/10.3858/emm.2010.42.2.013

Abstract

Influenza vaccine strains have been traditionally developed by annual reassortment between vaccine donor strain and the epidemic virulent strains. The classical method requires screening and genotyping of the vaccine strain among various reassortant viruses, which are usually laborious and time-consuming. Here we developed an efficient reverse genetic system to generate the 6:2 reassortant vaccine virus from cDNAs derived from the influenza RNAs. Thus, cDNAs of the two RNAs coding for surface antigens, haemagglutinin and neuraminidase from the epidemic virus and the 6 internal genes from the donor strain were transfected into cells and the infectious viruses of 6:2 defined RNA ratio were rescued. X-31 virus (a high-growth virus in embryonated eggs) and its cold-adapted strain X-31 ca were judiciously chosen as donor strains for the generation of inactivated vaccine and live-attenuated vaccine, respectively. The growth properties of these recombinant viruses in embryonated chicken eggs and MDCK cell were indistinguishable as compared to those generated by classical reassortment process. Based on the reverse genetic system, we generated 6 + 2 reassortant avian influenza vaccine strains corresponding to the A/Chicken/Korea/MS96 (H9N2) and A/Indonesia/5/2005 (H5N1). The results would serve as technical platform for the generation of both injectable inactivated vaccine and the nasal spray live attenuated vaccine for the prevention of influenza epidemics and pandemics.

Keyword

influenza vaccines; influenzavirus A; influenzavirus B; orthomyxoviridae

MeSH Terms

Animals
Chick Embryo
Chickens
Genetic Engineering
Hemagglutinins, Viral/genetics/metabolism
Humans
Influenza A Virus, H5N1 Subtype/*genetics/immunology
Influenza A Virus, H9N2 Subtype/*genetics/immunology
Influenza Vaccines/*genetics/metabolism
Influenza in Birds/immunology/virology
Influenza, Human/immunology/*prevention & control/virology
Neuraminidase/genetics/metabolism
Transgenes
Vaccines, Attenuated/*genetics/metabolism
Viral Proteins/genetics/metabolism