Hydrogen peroxide inactivation of influenza virus preserves antigenic structure and immunogenicity

J Virol Methods. 2014 Oct;207:232-7. doi: 10.1016/j.jviromet.2014.07.003. Epub 2014 Jul 12.


The use of live virus in the laboratory requires additional precautions, such as personnel training and special equipment, in order to limit the transmission risk. This is a requirement which not all laboratories can fulfill. In this study, a viral inactivation method is introduced using hydrogen peroxide (H2O2), which maintains antigenicity. Three strains of influenza viruses were inactivated and the ex vivo cellular and humoral immune responses were further analyzed, by comparing them to live viruses, in ELISpot, Multiplex and ELISA assays. In all assays, the H2O2 inactivated viruses displayed comparable responses to the live viruses, suggesting that the inactivated viruses still elicited immunogenic responses even though inactivation was confirmed by lack of viral replication in MDCK cells. Taken together, this study demonstrates that influenza viruses inactivated with H2O2 retain immunogenicity and are able to both detect humoral and elicit cellular immune responses in vitro, which could reduce the need to handle live viruses in the laboratory.

Keywords: Hydrogen peroxide; Immune response; Immunogenic; Inactivation; Influenza virus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Viral / drug effects*
  • Antigens, Viral / immunology*
  • Disinfectants / metabolism*
  • Dogs
  • Enzyme-Linked Immunosorbent Assay
  • Enzyme-Linked Immunospot Assay
  • Hydrogen Peroxide / metabolism*
  • Madin Darby Canine Kidney Cells
  • Microbial Viability / drug effects
  • Orthomyxoviridae / drug effects*
  • Orthomyxoviridae / immunology*
  • Orthomyxoviridae / physiology
  • Virus Cultivation
  • Virus Inactivation*
  • Virus Replication / drug effects


  • Antigens, Viral
  • Disinfectants
  • Hydrogen Peroxide