Comparative analysis of immunization schedules using a novel adenovirus-based immunotherapeutic targeting hepatitis B in naïve and tolerant mouse models

Vaccine. 2014 May 30;32(26):3256-63. doi: 10.1016/j.vaccine.2014.03.089. Epub 2014 Apr 13.


Development of active targeted immunotherapeutics is a rapid developing field in the arena of chronic infectious diseases. The question of repeated, closely spaced administration of immunotherapeutics to achieve a rapid impact on the replicating agent is an important one. We analyzed here, using a prototype adenovirus-based immunotherapeutic encoding Core and Polymerase from the hepatitis B virus (Ad-HBV), the influence of closely spaced repeated immunizations on the level and quality of induced HBV-specific and vector-specific immune responses in various mouse models. Ad-HBV, whether injected once or multiple times, was able to induce HBV- and adeno-specific T cells both in HBV-free mice and in a HBV tolerant mouse model. Adenovirus-specific T cell responses and titers of neutralizing anti-Ad5 antibodies increased from time of the 3rd injection. Interestingly, single or multiple Ad-HBV injections resulted in detection of Polymerase-specific functional T cells in HBV tolerant mice. Overall no modulation of the levels of HBV-specific cytokine-producing (IFNγ/TNFα) and cytolytic T cells was observed following repeated administrations (3 or 6 weekly injections) when compared with levels detected after a single injection with the exception of two markers: 1. the proportion of HBV-specific IFNγ-producing cells bearing the CD27+/CD43+ phenotype appeared to be sustained in C57BL/6J mice following 6 weekly injections; 2. the percentage of IFNγ/TNFα Core-specific producing cells observed in spleens of HLA-A2 mice as well as of that specific of Polymerase observed in livers of HBV tolerant mice was maintained. In addition, percentage of HBV-specific T cells expressing PD-1 was not increased by multiple injections. Overall these data show that, under experimental conditions used, rapid, closely spaced administrations of an adenovirus-based HBV immunotherapeutics does not inhibit induced T-cell responses including in a HBV-tolerant environment.

Keywords: Adenovirus; Hepatitis B virus; Immunotherapy; Multiple injections; T cells.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae
  • Animals
  • Gene Products, pol / immunology
  • HLA-A2 Antigen
  • Hepatitis B / prevention & control*
  • Hepatitis B Core Antigens / immunology
  • Hepatitis B Vaccines / immunology*
  • Immunity, Cellular*
  • Immunization Schedule*
  • Immunotherapy
  • Interferon-gamma / immunology
  • Liver / immunology
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Spleen / immunology
  • T-Lymphocytes / immunology
  • Tumor Necrosis Factor-alpha / immunology


  • Gene Products, pol
  • HLA-A*02:01 antigen
  • HLA-A2 Antigen
  • Hepatitis B Core Antigens
  • Hepatitis B Vaccines
  • P protein, Hepatitis B virus
  • Tumor Necrosis Factor-alpha
  • Interferon-gamma