Euroasian journal of hepato-gastroenterology

Register      Login

SEARCH WITHIN CONTENT

FIND ARTICLE

Volume / Issue

Online First

Archive
Volume  13 (2023)
Volume  12 (2022)
Volume  11 (2021)
Volume  10 (2020)
Volume  9 (2019)
Volume  8 (2018)
Volume  7 (2017)
Volume  6 (2016)
Volume  5 (2015)
Volume  4 (2014)
Volume  3 (2013)
Volume  2 (2012)
Volume  1 (2011)
Related articles

VOLUME 4 , ISSUE 2 ( July-December, 2014 ) > List of Articles

ORIGINAL ARTICLE

Optimization of a Therapeutic Vaccine Candidate by Studying Routes, Immunization Schedules and Antigen Doses in HBsAg-positive Transgenic Mice

H Trujillo, A Blanco, D García, F Freyre, J Aguiar, Y Lobaina, JC Aguilar

Citation Information : Trujillo H, Blanco A, García D, Freyre F, Aguiar J, Lobaina Y, Aguilar J. Optimization of a Therapeutic Vaccine Candidate by Studying Routes, Immunization Schedules and Antigen Doses in HBsAg-positive Transgenic Mice. Euroasian J Hepatogastroenterol 2014; 4 (2):70-78.

DOI: 10.5005/jp-journals-10018-1105

License: CC BY-NC 4.0

Published Online: 01-07-2013

Copyright Statement:  Copyright © 2014; The Author(s).


Abstract

Hepatitis B core antigens (HBcAg) and hepatitis B surface antigens (HBsAg) are the main structural antigens of hepatitis B virus (HBV). Both antigens are potent immunogens for experimental animals as well as in acutely infected patients. A novel formulation based on the combination of HBsAg and HBcAg has been developed as a therapeutic vaccine candidate, aimed at inducing an immune response capable of controlling the infection. An immunization schedule was conducted to evaluate the immunogenicity of this formulation after simultaneous immunization by the intranasal and parenteral routes using different schedules and doses. Humoral and cellular immune responses generated in blood and spleen were evaluated by engyme-linked immunosorbent assay (ELISA) and enzymeliked immunospot (ELISPOT) assays respectively. A first experiment evaluated two groups of mice simultaneously immunized by intranasal (IN) and subcutaneous (SC) routes, one including alum by SC route and, in the other, the formulation was injected without adjuvant. As a result, alum adjuvant did not increase the immunogenicity under the studied conditions. In fact, the group without alum induced the most potent immune response. The immune response was enhanced by combining IN and SC immunization compared to the SC route alone. In a second experiment, mice were immunized by different mucosal routes at the same time, and compared to the simultaneously (IN/SC) immunized groups. It was demonstrated that there is no improvement on the resulting immune response by using multiple routes of immunizations simultaneously; however, the increase of the antigen dose induced a superior immune response. Interestingly, the increase of antigen dose only by SC route did not favor the resulting immunogenicity. In conclusion, the use of HBsAg transgenic mice has proven useful to optimize the formulation, avoiding the unnecessary use of alum as adjuvant as well as provided information of the role of different mucosal immunization routes and antigen dose on the resulting immune response.

PDF Share
  1. Beasley RP. Hepatitis B virus. The major etiology for hepatocellular carcinoma. Cancer 1988 May;61(10):1942-1956
  2. Overview of the pathogenesis, prophylaxis and therapeusis of viral hepatitis B, with focus on reduction to practical applications. Vaccine 2001 Feb 28;19 (15-16):1837- 1848
  3. Hepatitis B virus and hepatitis delta virus. In: Mandell GL, Bennett JE, Dolin R, editors. Mandell, Douglas, and Bennett’s principles and practice of infectious diseases. 5th ed. New York: Elsevier/Churchill Livingstone; 2005. p. 1864-1890
  4. Seroconversion from hepatitis B e antigen to antibody in chronic type B hepatitis. Ann Intern Med 1981 Jan;94(6):744-748
  5. Immune response to hepatitis B virus core antigen (HBcAg): localization of T cell recognition sites within HBcAg/HBeAg. J Immunol 1987 Aug 15;139(4):1223-1231
  6. The nucleocapsid of hepatitis B virus is both a T-cell-independent and a T-cell-dependent antigen. Science 1986 Dec 12;234(4782):1398-1401
  7. Role of B cells in antigen presentation of hepatitis B core. Proc Natl Acad Sci USA 1997 Dec 23;94(26):14648-14653
  8. Molecular basis for the interaction of the hepatitis B virus core antigen with the surface immunoglobulin receptor on na€ive B cells. J Virol 2001;75(14):6367-6374
  9. Characterization of minor and major antigenic Table 2: Female vs male percentage of positive ELISPOT response in HBsAg-Tg mice immunized during the second experiment and HBsAg concentration in sera assessed at preimmune extraction Male Female Percentage of positive response by ELISPOT 1/9 (11%) 9/10 (90%) Conc. HBsAg (μg/ml) (time 0) 11.40 5.74 Table 1: Dynamic of positive ELISPOT response in HBsAg-Tg mice immunized with the formulation under study according to the schedule: HBs/HBcAg (IN) and HBs/HBcAg + Al(OH)3 (SC) during 10 administrations. Mice were immunized every 14 days, and spleen cells were isolated 10 days after the 3rd, 5th, 7th and 10th doses Doses and percentage of positive responses 3rd 5th 7th 10th HBs/HBcAg (IN) and HBs/HBcAg + Al(OH)3 (SC) 0/3 (0%) 1/3 (33%) 3/3 (100%) 3/3 (100%) H Trujillo et al 78 regions within the hepatitis B virus nucleocapsid. J Med Virol 1993 Nov;41(3):221-229
  10. Immunogenicity of peptide fusions to hepatitis B virus core antigen. Proc Natl Acad Sci USA 1989 Aug;86(16):6283-6287
  11. A molecular assembly system that renders antigens of choice highly repetitive for induction of protective B cell responses, Vaccine 2002 Aug 19;20(25-26):3104-3112
  12. Overview of the pathogenesis, prophylaxis and therapeusis of viral hepatitis B, with focus on reduction to practical applications. Vaccine 2001 Feb 28;19(15-16):1837-1848
  13. Multicenter study group. Efficacy and limitations of a specific immunotherapy in chronic hepatitis B. J Hepatol 2001 Jun;34(6):917-921
  14. In vivo immunization by vaccine therapy following virus suppression by lamivudine: a novel approach for treating patients with chronic hepatitis B. J Clin Virol 2005 Feb;32(2):156-161
  15. Therapeutic vaccination. Virus Res 2002 Jan 30;82(1-2):133-140
  16. Therapeutic vaccination of chronic hepatitis B patients with virus suppression by antiviral therapy: a randomized, controlled study of co-administration of HBsAg/ AS02 candidate vaccine and lamivudine. Vaccine 2007 Dec 12;25(51):8585-8597
  17. Quantity and quality of virus-specific CD8 cell response: relevance to the design of a therapeutic vaccine for chronic HBV infection. Mol Immunol 2001 Dec;38(6):467-473
  18. Translation of immunological knowledge into better treatments of chronic hepatitis B. J Hepatol 2003 Jul;39(1):115-124
  19. Development of a nasal vaccine for chronic hepatitis B infection that uses the ability of hepatitis B core antigen to stimulate a strong Th1 response against hepatitis B surface antigen. Immunol Cell Biol 2004 Oct;82(5):539-546
  20. Phase I clinical trial in healthy adults of a nasal vaccine candidate containing recombinant hepatitis B surface and core antigens. Int J Infect Dis 2007 Sep;11(5): 394-401
  21. Expresion del gen del antigeno de superficie del virus de la hepatitis B en ratones transgeicos. Interferón Biotecnología 1989;6:251-257
  22. Al. Characterization of Transgenic Mice Lineages II. Transgenic mice expressing HBsAg particles and showing female sterility. Acta Biotechnol 1993;13(4):373-383
  23. Mucosal immunogenicity of the hepatitis B core antigen. Biochem Biophys Res Commun 2003 Jan 17;300(3):745-750
  24. Similar as well as distinct MHC class I-binding peptides are generated by exogenous and endogenous processing of hepatitis B virus surface antigen. Eur J Immunol 1998 Dec;28(12):4149-4161
  25. DNA-mediated immunization in a transgenic mouse model of the hepatitis B surface antigen chronic carrier state. Proc Natl Acad Sci USA 1996 Oct 29;93(22): 12496-12501
  26. CpG Oligodeoxynucleotides with Hepatitis B surface antigen (HBsAg) for Vaccination in HBsAg-transgenic Mice. J Virol 2001 Jul;75(14):6482-6491
  27. Vaccine adjuvants revisited. Vaccine 2007 May 10;25(19):3752-3762
  28. Aluminium adjuvants-in retrospect and prospect. Vaccine 2004 Sep 9;22(27-28):3658-3668
  29. Aluminium compounds for use in vaccines. Immunol Cell Biol 2004 Oct;82(5):497-505
  30. The effect of the parenteral route of administration on the iImmune response to simultaneous nasal and parenteral immunizations using a new HBV therapeutic vaccine candidate. Viral Immunol 2010 Oct;23(5):521-529
  31. The nucleocapsid of hepatitis B virus is both a T-cell-independent and a T-cell-dependent antigen. Science 1986 Dec 12;234(4782):1398-1401
  32. Comparative study of the immunogenicity and immunoenhancing effects of two hepatitis B core antigen variantsin mice by nasal administration. Vaccine 2006 Apr 12;24(Suppl 2):S2-58-59
  33. Mucosal immunogenicity of the hepatitis B core antigen. Biochem Biophys Res Commun 2003 Jan 17;300(3):745-750
  34. Mechanism and therapeutic potential of DNA-based immunization against the envelope proteins of hepatitis B virus in normal and transgenic mice. Immunology 2001 May;103(1):90-97
  35. The nucleocapsid of the hepatitis B virus: a remarkable immunogenic structure. Antivir Res 2003 Oct;60(2):67-74
  36. Determinación del potencial irritante de un candidate vacunal nasal que combina al antígeno de superficie del virus de la hepatitis B con el antígeno de la nucleocápsida. Biotecnología Aplicada 2004; 21:143-147.
PDF Share
PDF Share

© Jaypee Brothers Medical Publishers (P) LTD.