Vaccine Design pp 221-244 | Cite as

Methods to Evaluate Novel Hepatitis C Virus Vaccines

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1403)

Abstract

The hepatitis C virus (HCV) is a major cause of severe liver disease worldwide. It is estimated that around 130–170 million individuals are chronic carriers of the infection and they are over time at an increased risk of developing severe liver disease. HCV is often referred to as a silent epidemic because the majority of infected individuals do not develop any symptoms. Hence, many individuals are diagnosed at a late stage and thus in need of immediate treatment. Today we have very effective direct-acting antivirals (DAAs), which cure more than 90–95 % of all treated patients. However, this treatment is associated with high-costs and the use is limited to the patients with most advanced liver disease in high-income countries. Notably, a majority of the chronic HCV carriers live in resource-poor countries and do not have access to the new effective DAAs. We therefore need to develop alternative treatments for chronic HCV infection such as therapeutic vaccines. The idea with therapeutic vaccines is to reactivate the infected patient’s own immune system. It is well known that patients with chronic HCV infection have dysfunctional immune responses to the virus. Hence, the vaccine should activate HCV-specific T cells that will home to the liver and eradicate the HCV infected hepatocytes. Importantly, one should also consider the combination of a therapeutic vaccine and DAAs as a treatment strategy to equip the resolving patients with post-cure HCV-specific immune responses. This would provide patients with a better protection against reinfection. Numerous genetic vaccine candidates for HCV have been developed and tested in clinical trials with limited effects on viral load and in general inefficient activation of HCV-specific immune responses. In this chapter we describe the rational of developing highly immunogenic vaccines for HCV. Different strategies to improve vaccine immunogenicity and methods to evaluate vaccine efficacy are described. Detailed description of vaccine delivery by intramuscular immunization in combination with in vivo electroporation/electrotransfer (EP/ET) is covered, as well as immunological analysis of primed immune responses by determination of interferon-γ (IFN-γ) production by ELISpot assay and direct ex vivo quantification of HCV NS3/4A-specific CD8+ T cells by pentamer staining. To analyze the in vivo functionality of primed NS3/4A-specific T cells we utilized the in vivo bioluminescence imaging technology. In conclusion, this chapter describes a method to design HCV vaccines and also a protocol to assess their efficacy.

Key words

Hepatitis C virus HCV Vaccine Genetic vaccination Delivery ELISpot Pentamer staining In vivo imaging 

Notes

Acknowledgements

The following work was supported by grants from the Swedish Research Council (K2012-99X-22017-01-3), the Swedish Society of Medicine, Goljes Memorial Fund, the Åke Wiberg Foundation, the Ruth and Richard Juhlin Foundation, and from Karolinska Institutet.

References

  1. 1.
    Mohd Hanafiah K, Groeger J, Flaxman AD, Wiersma ST (2013) Global epidemiology of hepatitis C virus infection: new estimates of age-specific antibody to HCV seroprevalence. Hepatology 57:1333–1342CrossRefPubMedGoogle Scholar
  2. 2.
    Thomas DL (2013) Global control of hepatitis C: where challenge meets opportunity. Nature Med 19:850–858CrossRefPubMedGoogle Scholar
  3. 3.
    Sulkowski MS, Gardiner DF, Rodriguez-Torres M, Reddy KR et al (2014) Daclatasvir plus sofosbuvir for previously treated or untreated chronic HCV infection. New Engl J Med 370:211–221CrossRefPubMedGoogle Scholar
  4. 4.
    Feld JJ, Kowdley KV, Coakley E, Sigal S et al (2014) Treatment of HCV with ABT-450/r–ombitasvir and dasabuvir with ribavirin. New Engl J Med 370:1594–1603CrossRefPubMedGoogle Scholar
  5. 5.
    Ahlen G, Frelin L, Brenndorfer ED, Brass A et al (2013) Containing “The Great Houdini” of viruses: combining direct acting antivirals with the host immune response for the treatment of chronic hepatitis C. Drug Resist Updat 16:60–67CrossRefPubMedGoogle Scholar
  6. 6.
    Callendret B, Eccleston HB, Hall S, Satterfield W et al (2014) T-cell immunity and hepatitis C virus reinfection after cure of chronic hepatitis C with an interferon-free antiviral regimen in a chimpanzee. Hepatology 60:1531–1540CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Barnes E, Folgori A, Capone S, Swadling L et al (2012) Novel adenovirus-based vaccines induce broad and sustained T cell responses to HCV in man. Sci Transl Med 4:115ra111CrossRefGoogle Scholar
  8. 8.
    Swadling L, Capone S, Antrobus RD, Brown A et al (2014) A human vaccine strategy based on chimpanzee adenoviral and MVA vectors that primes, boosts, and sustains functional HCV-specific T cell memory. Sci Transl Med 6:261ra153CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Weiland O, Ahlen G, Diepolder H, Jung MC et al (2013) Therapeutic DNA vaccination using in vivo electroporation followed by standard of care therapy in patients with genotype 1 chronic hepatitis C. Mol Therapy 21:1796–1805CrossRefGoogle Scholar
  10. 10.
    Di Bisceglie AM, Janczweska-Kazek E, Habersetzer F et al (2014) Efficacy of immunotherapy with TG4040, peg-interferon, and ribavirin in a Phase 2 study of patients with chronic HCV infection. Gastroenterology 147:119–131CrossRefPubMedGoogle Scholar
  11. 11.
    Habersetzer F, Honnet G, Bain C, Maynard-Muet M et al (2011) A poxvirus vaccine is safe, induces T-cell responses, and decreases viral load in patients with chronic hepatitis C. Gastroenterology 141:890–899CrossRefPubMedGoogle Scholar
  12. 12.
    Klade CS, Schuller E, Boehm T, von Gabain A, Manns MP (2012) Sustained viral load reduction in treatment-naive HCV genotype 1 infected patients after therapeutic peptide vaccination. Vaccine 30:2943–2950CrossRefPubMedGoogle Scholar
  13. 13.
    Wedemeyer H, Schuller E, Schlaphoff V, Stauber RE et al (2009) Therapeutic vaccine IC41 as late add-on to standard treatment in patients with chronic hepatitis C. Vaccine 27:5142–5151CrossRefPubMedGoogle Scholar
  14. 14.
    Frelin L, Ahlen G, Alheim M, Weiland O et al (2004) Codon optimization and mRNA amplification effectively enhances the immunogenicity of the hepatitis C virus nonstructural 3/4A gene. Gene Ther 11:522–533CrossRefPubMedGoogle Scholar
  15. 15.
    Chen A, Ahlen G, Brenndorfer ED, Brass A et al (2011) Heterologous T cells can help restore function in dysfunctional hepatitis C virus nonstructural 3/4A-specific T cells during therapeutic vaccination. J Immunol 186:5107–5118CrossRefPubMedGoogle Scholar
  16. 16.
    Levander S, Sällberg M, Ahlén G, Frelin L. A non-human hepadnaviral adjuvant for hepatitis C virus-based DNA vaccines. Submitted for publication.Google Scholar
  17. 17.
    Brass A, Frelin L, Milich DR, Sallberg M, Ahlen G (2015) Functional aspects of intrahepatic Hepatitis B Virus-specific T cells induced by therapeutic DNA vaccination. Mol Therapy 23:578–590CrossRefGoogle Scholar
  18. 18.
    Tovey MG, Lallemand C (2010) Adjuvant activity of cytokines. Methods Mol Biol 626:287–309CrossRefPubMedGoogle Scholar
  19. 19.
    Applequist SE, Rollman E, Wareing MD, Liden M et al (2005) Activation of innate immunity, inflammation, and potentiation of DNA vaccination through mammalian expression of the TLR5 agonist flagellin. J Immunol 175:3882–3891CrossRefPubMedGoogle Scholar
  20. 20.
    Branden LJ, Mohamed AJ, Smith CI (1999) A peptide nucleic acid-nuclear localization signal fusion that mediates nuclear transport of DNA. Nature Biotechnol 17:784–787CrossRefGoogle Scholar
  21. 21.
    Sallberg M, Frelin L, Ahlen G, Sallberg-Chen M (2015) Electroporation for therapeutic DNA vaccination in patients. Medical Microbiol Immunol 204:131–135CrossRefGoogle Scholar
  22. 22.
    Ulmer JB, Donnelly JJ, Parker SE, Rhodes GH et al (1993) Heterologous protection against influenza by injection of DNA encoding a viral protein. Science 259:1745–1749CrossRefPubMedGoogle Scholar
  23. 23.
    MacGregor RR, Boyer JD, Ugen KE, Lacy KE et al (1998) First human trial of a DNA-based vaccine for treatment of human immunodeficiency virus type 1 infection: safety and host response. J Infect Dis 178:92–100CrossRefPubMedGoogle Scholar
  24. 24.
    Calarota S, Bratt G, Nordlund S, Hinkula J et al (1998) Cellular cytotoxic response induced by DNA vaccination in HIV-1-infected patients. Lancet 351:1320–1325CrossRefPubMedGoogle Scholar
  25. 25.
    Aihara H, Miyazaki J (1998) Gene transfer into muscle by electroporation in vivo. Nature Biotechnol 16:867–870CrossRefGoogle Scholar
  26. 26.
    Ahlen G, Soderholm J, Tjelle T, Kjeken R, Frelin L et al (2007) In vivo electroporation enhances the immunogenicity of hepatitis C virus nonstructural 3/4A DNA by increased local DNA uptake, protein expression, inflammation, and infiltration of CD3+ T cells. J Immunol 179:4741–4753CrossRefPubMedGoogle Scholar
  27. 27.
    Gothelf A, Gehl J (2012) What you always needed to know about electroporation based DNA vaccines. Human Vaccines Immunother 8:1694–1702CrossRefGoogle Scholar
  28. 28.
    Ahlen G, Sallberg M, Frelin L (2013) Methods for monitoring gene gun-induced HBV- and HCV-specific immune responses in mouse models. Methods Mol Biol 940:239–267PubMedGoogle Scholar
  29. 29.
    Fournillier A, FrelinL JE, Ahlen G et al (2013) A heterologous prime/boost vaccination strategy enhances the immunogenicity of therapeutic vaccines for hepatitis C virus. J Infect Dis 208:1008–1019CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Lin Y, Kwon T, Polo J, Zhu YF et al (2008) Induction of broad CD4+ and CD8+ T-cell responses and cross-neutralizing antibodies against hepatitis C virus by vaccination with Th1-adjuvanted polypeptides followed by defective alphaviral particles expressing envelope glycoproteins gpE1 and gpE2 and nonstructural proteins 3, 4, and 5. J Virol 82:7492–7503CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Pancholi P, Perkus M, Tricoche N, Liu Q, Prince AM (2003) DNA immunization with hepatitis C virus (HCV) polycistronic genes or immunization by HCV DNA priming-recombinant canarypox virus boosting induces immune responses and protection from recombinant HCV-vaccinia virus infection in HLA-A2.1-transgenic mice. J Virol 77:382–390CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Laboratory Medicine, Division of Clinical Microbiology, F68, Karolinska InstitutetKarolinska University Hospital HuddingeStockholmSweden

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