Genetic engineering of live attenuated influenza viruses

Methods Mol Biol. 2012:865:163-74. doi: 10.1007/978-1-61779-621-0_10.

Abstract

The first live attenuated influenza vaccine (LAIV) was licensed in the USA in 2003; it is a trivalent vaccine composed of two type A (H1N1 and H3N2) and one type B influenza virus each at 10(7) fluorescent focus units (FFU). Each influenza vaccine strain is a reassortant virus that contains the hemagglutinin (HA) and neuraminidase (NA) gene segments from a wild-type influenza virus and the six internal protein gene segments from a master donor virus (MDV) of either cold-adapted A/Ann Arbor/6/60 or B/Ann Arbor/1/66. MDV confers the cold-adapted, temperature-sensitive, and attenuation phenotypes to the vaccine strains. The reassortant vaccine seeds are currently produced by reverse genetics and amplified in specific pathogen-free (SPF) 9-11 days old embryonated chicken eggs for manufacture. In addition, MDCK cell culture manufacture processes have been developed to produce LAIV for research use and with modifications for clinical and/or commercial grade material production.

MeSH terms

  • Animals
  • Cell Line
  • Chick Embryo
  • Cold Temperature
  • Culture Media, Serum-Free
  • Dogs
  • Genetic Engineering / methods
  • Hemagglutinins / genetics*
  • Hemagglutinins / immunology
  • Humans
  • Influenza Vaccines / biosynthesis*
  • Influenza Vaccines / genetics
  • Influenza Vaccines / immunology
  • Influenza, Human / immunology
  • Influenza, Human / prevention & control*
  • Influenza, Human / virology
  • Neuraminidase / genetics*
  • Neuraminidase / immunology
  • Orthomyxoviridae / genetics*
  • Orthomyxoviridae / immunology
  • Plasmids
  • Reassortant Viruses / genetics*
  • Reassortant Viruses / immunology
  • Reverse Genetics / methods
  • Vaccines, Attenuated
  • Viral Proteins / genetics*
  • Viral Proteins / immunology
  • Virus Replication

Substances

  • Culture Media, Serum-Free
  • Hemagglutinins
  • Influenza Vaccines
  • Vaccines, Attenuated
  • Viral Proteins
  • Neuraminidase