Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Synthetic DNA immunogen encoding hepatitis B core antigen drives immune response in liver

Cancer Gene Therapy volume 19, pages 779–787 (2012)Cite this article

Subjects

Abstract

The prevalence of hepatitis B virus (HBV) infection in Asia and sub-Sahara Africa is alarming. With quarter of a billion people chronically infected worldwide and at risk of developing liver cancer, the need for a prophylactic or therapeutic vaccination approach that can effectively induce protective responses against the different genotypes of HBV is more important than ever. Such a strategy will require both the induction of a strong antigen-specific immune response and the subsequent deployment of immune response towards the liver. Here, we assessed the ability of a synthetic DNA vaccine encoding a recombinant consensus plasmid from genotype A through E of the HBV core antigen (HBcAg), to drive immunity in the liver. Intramuscular vaccination induced both strong antigen-specific T cell and high titer antibody responses systematically and in the liver. Furthermore, immunized mice showed strong cytotoxic responses that eliminate adoptively transferred HBV-coated target cells. Importantly, vaccine-induced immune responses provided protection from HBcAg plasmid-based liver transfection in a hydrodynamic liver transfection model. These data provide important insight into the generation of peripheral immune responses that are recruited to the liver—an approach that can be beneficial in the search for vaccines or immune-therapies to liver disease.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

Abbreviations

CTL:

cytotoxic T lymphocyte

EP:

electroporation

HA:

hemagglutinin

HBcAg:

hepatitis B virus core antigen

IFN-γ:

interferon-gamma

PBS:

phosphate-buffered saline

TNF-α:

tumor necrosis factor-alpha

References

  1. Parkin DM, Bray F, Ferlay J, Pisani P . Estimating the world cancer burden: Globocan 2000. Int J Cancer 2001; 94: 153–156.

    Article  CAS  Google Scholar 

  2. El-Serag HB, Mason AC . Rising incidence of hepatocellular carcinoma in the United States. N Engl J Med 1999; 340: 745–750.

    Article  CAS  Google Scholar 

  3. Kurbanov F, Tanaka Y, Mizokami M . Geographical and genetic diversity of the human hepatitis B virus. Hepatol Res 2010; 40: 14–30.

    Article  Google Scholar 

  4. Williams R . Global challenges in liver disease. Hepatology 2006; 44: 521–526.

    Article  Google Scholar 

  5. Shepard CW, Simard EP, Finelli L, Fiore AE, Bell BP . Hepatitis B virus infection: epidemiology and vaccination. Epidemiol Rev 2006; 28: 112–125.

    Article  Google Scholar 

  6. Michel ML, Mancini-Bourgine M . Therapeutic vaccination against chronic hepatitis B virus infection. J Clin Virol 2005; 34 (Supplement 1): S108–S114.

    Article  CAS  Google Scholar 

  7. Cao G-W . Clinical relevance and public health significance of hepatitis B virus genomic variations. World J Gastroenterol 2009; 15: 5761–5769.

    Article  CAS  Google Scholar 

  8. Lin C-L, Kao J-H . The clinical implications of hepatitis B virus genotype: recent advances. J Gastroenterol Hepatol 2011; 26: 123–130.

    Article  Google Scholar 

  9. Torresi J . The virological and clinical significance of mutations in the overlapping envelope and polymerase genes of hepatitis B virus. J Clin Virol 2002; 25: 97–106.

    Article  CAS  Google Scholar 

  10. Yan J, Corbitt N, Pankhong P, Shin T, Khan A, Sardesai NY et al. Immunogenicity of a novel engineered HIV-1 clade C synthetic consensus-based envelope DNA vaccine. Vaccine 2011; 29: 7173–7181.

    Article  CAS  Google Scholar 

  11. Lin Y-J, Huang L-R, Yang H-C, Tzeng H-T, Hsu P-N, Wu H-L et al. Hepatitis B virus core antigen determines viral persistence in a C57BL/6 mouse model. Proc Natl Acad Sci 2010; 107: 9340–9345.

    Article  CAS  Google Scholar 

  12. Ferrari C, Penna A, Bertoletti A, Valli A, Antoni AD, Giuberti T et al. Cellular immune response to hepatitis B virus-encoded antigens in acute and chronic hepatitis B virus infection. J Immunol 1990; 145 (10): 3442–3449.

    CAS  PubMed  Google Scholar 

  13. Tsai SL, Chen PJ, Lai MY, Yang PM, Sung JL, Huang JH et al. Acute exacerbations of chronic type B hepatitis are accompanied by increased T cell responses to hepatitis B core and e antigens. Implications for hepatitis B e antigen seroconversion. J Clin Invest 1992; 89 (1): 87–96.

    Article  CAS  Google Scholar 

  14. Xing Y-P, Huang Z-H, Wang S-X, Cai J, Li J, Chou T-H et al. Novel DNA vaccine based on hepatitis B virus core gene induces specific immune responses in Balb/c mice. World J Gastroenterol 2005; 11 (29): 4583–4586.

    Article  CAS  Google Scholar 

  15. Hirao LA, Wu L, Khan AS, Satishchandran A, Draghia-Akli R, Weiner DB . Intradermal/subcutaneous immunization by electroporation improves plasmid vaccine delivery and potency in pigs and rhesus macaques. Vaccine 2008; 26 (3): 440–448.

    Article  CAS  Google Scholar 

  16. Laddy DJ, Yan J, Kutzler M, Kobasa D, Kobinger GP, Khan AS et al. Heterosubtypic protection against pathogenic human and avian influenza viruses via in vivo electroporation of synthetic consensus DNA antigens. PLoS ONE 2008; 3 (6): e2517.

    Article  Google Scholar 

  17. Yan J, Harris K, Khan AS, Draghia-Akli R, Sewell D, Weiner DB . Cellular immunity induced by a novel HPV18 DNA vaccine encoding an E6/E7 fusion consensus protein in mice and rhesus macaques. Vaccine 2008; 26 (40): 5210–5215.

    Article  CAS  Google Scholar 

  18. Hirao LA, Wu L, Khan AS, Hokey DA, Yan J, Dai A et al. Combined effects of IL-12 and electroporation enhances the potency of DNA vaccination in macaques. Vaccine 2008; 26 (25): 3112–3120.

    Article  CAS  Google Scholar 

  19. Shedlock DJ, Talbott KT, Cress C, Ferraro B, Tuyishme S, Mallilankaraman K et al. A highly optimized DNA vaccine confers complete protective immunity against high-dose lethal lymphocytic choriomeningitis virus challenge. Vaccine 2011; 29: 6755–6762.

    Article  CAS  Google Scholar 

  20. Choo DK, Murali-Krishna K, Anita R, Ahmed R . Homeostatic turnover of virus-specific memory CD8 T cells occurs stochastically and is independent of CD4 T cell help. J Immunol 2010; 185 (6): 3436–3444.

    Article  CAS  Google Scholar 

  21. Kutzler MA, Robinson TM, Chattergoon MA, Choo DK, Choo AY, Choe PY et al. Coimmunization with an optimized IL-15 plasmid results in enhanced function and longevity of CD8 T cells that are partially independent of CD4 T cell help. J Immunol 2005; 175 (1): 112–123.

    Article  CAS  Google Scholar 

  22. Shedlock DJ, Talbott KT, Morrow MP, Ferraro B, Hokey DA, Muthumani K et al. Ki-67 staining for determination of rhesus macaque T cell proliferative responses ex vivo. Cytometry Part A 2010; 77A (3): 275–284.

    Google Scholar 

  23. Barber DL, Wherry EJ, Ahmed R . Cutting edge: rapid in vivo killing by memory CD8 T cells. J Immunol 2003; 171 (1): 27–31.

    Article  CAS  Google Scholar 

  24. Durward MA, Harms J, Magnani DM, Eskra L, Splitter GA . Discordant brucella melitensis antigens yield cognate CD8+ T cells in vivo. Infect Immun 2010; 78 (1): 168–176.

    Article  CAS  Google Scholar 

  25. Zhang G, Budker V, Wolff JA . High levels of foreign gene expression in hepatocytes after tail vein injections of naked plasmid DNA. Hum Gene Ther 1999; 10 (10): 1735–1737.

    Article  CAS  Google Scholar 

  26. Ahlen G, Nystrom J, Pult I, Frelin L, Hultgren C, Sallberg M . In vivo clearance of hepatitis C virus nonstructural 3/4A-expressing hepatocytes by DNA vaccine-primed cytotoxic T lymphocytes. J Infect Dis 2005; 192 (12): 2112–2116.

    Article  Google Scholar 

  27. Yang PL, Althage A, Chung J, Chisari FV . Hydrodynamic injection of viral DNA: a mouse model of acute hepatitis B virus infection. Proc Natl Acad Sci 2002; 99 (21): 13825–13830.

    Article  CAS  Google Scholar 

  28. Penna A, Artini M, Cavalli A, Levrero M, Bertoletti A, Pilli M et al. Long-lasting memory T cell responses following self-limited acute hepatitis B. J Clin Invest 1996; 98 (5): 1185–1194.

    Article  CAS  Google Scholar 

  29. Thimme R, Wieland S, Steiger C, Ghrayeb J, Reimann KA, Purcell RH et al. CD8+ T cells mediate viral clearance and disease pathogenesis during acute hepatitis B virus infection. J Virol 2003; 77 (1): 68–76.

    Article  CAS  Google Scholar 

  30. Guidotti LG, Ishikawa T, Hobbs MV, Matzke B, Schreiber R, Chisari FV . Intracellular inactivation of the hepatitis B virus by cytotoxic T lymphocytes. Immunity 1996; 4 (1): 25–36.

    Article  CAS  Google Scholar 

  31. Biermer M, Puro R, Schneider RJ . Tumor necrosis factor alpha inhibition of hepatitis B virus replication involves disruption of capsid integrity through activation of NF-{kappa}B. J Virol 2003; 77 (7): 4033–4042.

    Article  CAS  Google Scholar 

  32. Chang JJ, Wightman F, Bartholomeusz A, Ayres A, Kent SJ, Sasadeusz J et al. Reduced hepatitis B virus (HBV)-specific CD4+ T-cell responses in human immunodeficiency virus type 1-HBV-coinfected individuals receiving HBV-active antiretroviral therapy. J Virol 2005; 79 (5): 3038–3051.

    Article  CAS  Google Scholar 

  33. Reignat S, Webster GJM, Brown D, Ogg GS, King A, Seneviratne SL et al. Escaping high viral load exhaustion. J Exp Med 2002; 195 (9): 1089–1101.

    Article  CAS  Google Scholar 

  34. Chang JJL, Sharon R . Immunopathogenesis of hepatitis B virus infection. Immunol Cell Biol 2006; 85: 1.

    Google Scholar 

  35. Pillet SP, Kobasa D, Meunier I, Gray M, Laddy D, Weiner DB et al. Cellular immune response in the presence of protective antibody levels correlates with protection against 1918 influenza in ferrets. Vaccine 2011; 29: 6793–6801.

    Article  CAS  Google Scholar 

  36. Peng J-L, Zhao Y-G, Mai J-H, Pang W-K, Guo W, Chen G-M et al. Non-cytolytic antigen clearance in DNA-vaccinated mice with electroporation. Acta Pharmacol Sin 2007; 28 (7): 1024–1030.

    Article  CAS  Google Scholar 

  37. Kuhröber A, Wild J, Pudollek HP, Chisari FV, Reimann J . DNA vaccination with plasmids encoding the intracellular (HBcAg) or secreted (HBeAg) form of the core protein of hepatitis B virus primes T cell responses to two overlapping Kb- and Kd-restricted epitopes. Int Immunol 1997; 9 (8): 1203–1212.

    Article  Google Scholar 

  38. Sällberg M, Hughes J, Javadian A, Ronlov G, Hultgren C, Townsend K et al. Genetic immunization of chimpanzees chronically infected with the hepatitis B virus, using a recombinant retroviral vector encoding the hepatitis B virus core antigen. Hum Gene Ther 1998; 9 (12): 1719–1729.

    Article  Google Scholar 

  39. Livingston BD, Crimi C, Fikes J, Chesnut RW, Sidney J, Sette A . Immunization with the HBV core 18-27 epitope elicits CTL responses in humans expressing different HLA-A2 supertype molecules. Hum Immunol 1999; 60 (11): 1013–1017.

    Article  CAS  Google Scholar 

  40. Kannanganat S, Ibegbu C, Chennareddi L, Robinson HL, Amara RR . Multiple-cytokine-producing antiviral CD4 T cells are functionally superior to single-cytokine-producing cells. J Virol 2007; 81 (16): 8468–8476.

    Article  CAS  Google Scholar 

  41. Yin Y, Wu C, Song J, Wang J, Zhang E, Liu H et al. DNA immunization with fusion of CTLA-4 to hepatitis B virus (HBV) core protein enhanced Th2 type responses and cleared HBV with an accelerated kinetic. PLoS ONE 2011; 6 (7): e22524.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    N Obeng-Adjei, D K Choo, J Saini, P Pankhong, A Parikh, J S Chu & D B Weiner

  2. Inovio Pharmaceuticals Inc, 1787 Sentry Parkway West, Blue Bell, PA, USA

    J Yan

Authors
  1. N Obeng-Adjei
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. D K Choo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J Saini
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J Yan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. P Pankhong
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. A Parikh
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. J S Chu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. D B Weiner
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to D B Weiner.

Ethics declarations

Competing interests

DBW and his laboratory have several commercial relationships. He has received consulting fees, stock ownership, for Advisory Board/Review Board Service, speaking support and research support from commercial entities, including Inovio, BMS, VGXI, Pfizer, Virxsys, J & J, Merck, Sanofi Pasteur, Althea, Novo Nordisk, SSI, Aldevron, Novartis, Incyte and possibly others. No writing assistance was utilized in the production of this manuscript. The other authors declare no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Obeng-Adjei, N., Choo, D., Saini, J. et al. Synthetic DNA immunogen encoding hepatitis B core antigen drives immune response in liver. Cancer Gene Ther 19, 779–787 (2012). https://doi.org/10.1038/cgt.2012.61

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/cgt.2012.61

Keywords

This article is cited by

Search

Advanced search

Quick links