Flavivirus DNA Vaccine Design and Adjuvant Selection

  • Lei Li
  • Yoshikazu Honda-Okubo
  • Nikolai PetrovskyEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2183)


A DNA vaccine is a plasmid encoding a vaccine antigen together with an efficient eukaryotic promoter to drive protein expression. The chief problem of DNA vaccines has been their suboptimal immunogenicity in humans. Many different flaviviruses infect and cause serious illness and even death in humans, but human vaccines are not available against most of the relevant flaviviruses with the exception of Japanese encephalitis virus. DNA vaccines are easy and fast to produce at relatively low cost, do not require handling of dangerous pathogens, are stable at room temperature allowing for low-cost storage and transportation, and are highly versatile, allowing for rapid changes in coding sequence design and synthesis. This makes a DNA vaccine approach ideally suited for development as a broad-based flavivirus vaccine platform. However, to be useful as a flavivirus prophylactic vaccine platform in humans, a method would need to be found to enhance DNA vaccine immunogenicity without the need for the cumbersome and expensive equipment involved with electroporation. We describe here a protocol used to test different adjuvants with flavivirus DNA vaccines to determine an optimal formulation. An optimal regimen involving a DNA adjuvanted vaccine prime followed by an adjuvanted protein vaccine boost is described and can be applied by readers to solve barriers to the development of other DNA vaccines where immunogenicity is a problem.

Key words

DNA vaccine Flavivirus Adjuvant Codon optimization Electroporation 



This work was supported by National Institute of Allergy and Infectious Diseases of the National Institutes of Health under Contracts No. HHS-N272200800039C and HHS-N272201400053C.


  1. 1.
    Li L, Petrovsky N (2017) Molecular adjuvants for DNA vaccines. Curr Issues Mol Biol 22:17–40CrossRefGoogle Scholar
  2. 2.
    Li L, Saade F, Petrovsky N (2012) The future of human DNA vaccines. J Biotechnol 162(2–3):171–182CrossRefGoogle Scholar
  3. 3.
    Porter KR et al (2011) Immunogenicity and protective efficacy of a vaxfectin-adjuvanted tetravalent dengue DNA vaccine. Vaccine 30(2):336–341CrossRefGoogle Scholar
  4. 4.
    Nukuzuma C et al (2003) Enhancing effect of vaxfectin on the ability of a Japanese encephalitis DNA vaccine to induce neutralizing antibody in mice. Viral Immunol 16(2):183–189CrossRefGoogle Scholar
  5. 5.
    Ara Y et al (2001) Zymosan enhances the immune response to DNA vaccine for human immunodeficiency virus type-1 through the activation of complement system. Immunology 103(1):98–105CrossRefGoogle Scholar
  6. 6.
    Honda-Okubo Y, Saade F, Petrovsky N (2012) Advax, a polysaccharide adjuvant derived from delta inulin, provides improved influenza vaccine protection through broad-based enhancement of adaptive immune responses. Vaccine 30(36):5373–5381CrossRefGoogle Scholar
  7. 7.
    Lobigs M et al (2010) An inactivated Vero cell-grown Japanese encephalitis vaccine formulated with Advax, a novel inulin-based adjuvant, induces protective neutralizing antibody against homologous and heterologous flaviviruses. J Gen Virol 91(Pt 6):1407–1417CrossRefGoogle Scholar
  8. 8.
    Cukjati D et al (2007) Real time electroporation control for accurate and safe in vivo non-viral gene therapy. Bioelectrochemistry 70(2):501–507CrossRefGoogle Scholar
  9. 9.
    Becker SM, Kuznetsov AV (2007) Local temperature rises influence in vivo electroporation pore development: a numerical stratum corneum lipid phase transition model. J Biomech Eng 129(5):712–721CrossRefGoogle Scholar
  10. 10.
    Wu CJ et al (2004) In vivo electroporation of skeletal muscles increases the efficacy of Japanese encephalitis virus DNA vaccine. Vaccine 22(11–12):1457–1464CrossRefGoogle Scholar
  11. 11.
    Livingston BD et al (2009) Comparative performance of a licensed anthrax vaccine versus electroporation based delivery of a PA encoding DNA vaccine in rhesus macaques. Vaccine 28(4):1056–1061CrossRefGoogle Scholar
  12. 12.
    Luckay A et al (2007) Effect of plasmid DNA vaccine design and in vivo electroporation on the resulting vaccine-specific immune responses in rhesus macaques. J Virol 81(10):5257–5269CrossRefGoogle Scholar
  13. 13.
    Simon AJ et al (2008) Enhanced in vivo transgene expression and immunogenicity from plasmid vectors following electrostimulation in rodents and primates. Vaccine 26(40):5202–5209CrossRefGoogle Scholar
  14. 14.
    Schneeweiss A et al (2011) A DNA vaccine encoding the E protein of West Nile virus is protective and can be boosted by recombinant domain DIII. Vaccine 29(37):6352–6357CrossRefGoogle Scholar
  15. 15.
    Li P et al (2008) Enhancement of humoral and cellular immunity in mice against Japanese encephalitis virus using a DNA prime - protein boost vaccine strategy. Vet J 183(2):210–216CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  • Lei Li
    • 1
    • 2
  • Yoshikazu Honda-Okubo
    • 1
    • 2
  • Nikolai Petrovsky
    • 1
    • 2
    Email author
  1. 1.Vaxine Pty LtdWarradaleAustralia
  2. 2.College of Medicine and Public HealthFlinders UniversityAdelaideAustralia

Personalised recommendations