Polyvalent DNA vaccines with bidirectional promoters

The hepatitis B surface antigen (HBsAg) and core antigen (HBcAg) were coexpressed from a synthetic bidirectional promoter with the tetracycline-inactivated transactivator (tTA). The function of this autoregulative system was evaluated following either transfer into established cell lines or intramuscular and intradermal injection of high or low doses of DNA into mice. We measured in vitro antigen expression and in vivo the induction of specific humoral and cellular immune responses. Successful regulation of antigen expression was observed in cultured cells. DNA vaccination with these constructs efficiently primed hepatitis B virus (HBV) specific immunity. However, immunogenic concentrations of the antigens were expressed even in the absence of the transactivator, indicating that low expression level is sufficient to prime an immune response. The bidirectional promoter allows coexpression of either both HBV antigens or a HBV antigen and enhanced green fluorescent protein leading to efficient priming of stable immunity against both antigens. This study demonstrates the potential of synthetic polyvalent plasmids in DNA vaccination.

[1]  M. Rivkina,et al.  Costimulatory protein B7-1 enhances the cytotoxic T cell response and antibody response to hepatitis B surface antigen. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[2]  U. Certa,et al.  Efficient control of gene expression by single step integration of the tetracycline system in transgenic mice , 1996, Nature Biotechnology.

[3]  D. Schatz,et al.  A modified tetracycline-regulated system provides autoregulatory, inducible gene expression in cultured cells and transgenic mice. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[4]  H. Blau,et al.  Tetracycline-regulated gene expression following direct gene transfer into mouse skeletal muscle , 1995, Somatic cell and molecular genetics.

[5]  M. Schleef,et al.  DNA-mediated immunization to the hepatitis B surface antigen in mice: aspects of the humoral response mimic hepatitis B viral infection in humans. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[6]  J. Reimann,et al.  Processing of exogenous heat-aggregated (denatured) and particulate (native) hepatitis B surface antigen for class I-restricted epitope presentation. , 1995, Journal of immunology.

[7]  F. Chisari,et al.  Nucleic acid vaccination primes hepatitis B , 1995 .

[8]  J. Wands,et al.  Cytokine and hepatitis B virus DNA Co‐immunizations enhance cellular and humoral immune responses to the middle but not to the large hepatitis B virus surface antigen in mice , 1998, Hepatology.

[9]  H. Hauser,et al.  Polyvalent vaccination against hepatitis B surface and core antigen using a dicistronic expression plasmid. , 1998, Vaccine.

[10]  M. Gossen,et al.  Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[11]  M. Gossen,et al.  Co-regulation of two gene activities by tetracycline via a bidirectional promoter. , 1995, Nucleic acids research.

[12]  F. Chisari,et al.  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. , 1997, International immunology.

[13]  H. Davis,et al.  DNA-based immunization induces continuous secretion of hepatitis B surface antigen and high levels of circulating antibody. , 1993, Human molecular genetics.

[14]  S. Prusiner,et al.  Doxycycline control of prion protein transgene expression modulates prion disease in mice. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[15]  V. Zurawski,et al.  Cellular and humoral immune response to hepatitis B virus structural proteins in mice after DNA-based immunization. , 1997, Gastroenterology.

[16]  M. Gossen,et al.  Doxycycline-mediated quantitative and tissue-specific control of gene expression in transgenic mice. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[17]  F. Chisari,et al.  Hepatitis B virus immunopathogenesis. , 1995, Annual review of immunology.

[18]  A. Magalini,et al.  Specificity of action of a herpes virus VP16/tetracycline-dependent trans-activator in mammalian cell cultures. , 1995, DNA and cell biology.

[19]  M. Gossen,et al.  Tetracycline-controlled transcription in eukaryotes: novel transactivators with graded transactivation potential. , 1997, Nucleic acids research.

[20]  F. Chisari,et al.  Polyclonality and multispecificity of the CTL response to a single viral epitope. , 1998, Journal of immunology.

[21]  F. Chisari,et al.  DNA immunization induces antibody and cytotoxic T cell responses to hepatitis B core antigen in H-2b mice. , 1996, Journal of immunology.

[22]  Y. Barenholz,et al.  Exogenous hepatitis B surface antigen particles processed by dendritic cells or macrophages prime murine MHC class I-restricted cytotoxic T lymphocytes in vivo. , 1995, Journal of immunology.

[23]  J. Reimann,et al.  DNA-mediated immunization in mice induces a potent MHC class I-restricted cytotoxic T lymphocyte response to the hepatitis B envelope protein. , 1995, Human gene therapy.

[24]  J. Reimann,et al.  DNA vector constructs that prime hepatitis B surface antigen-specific cytotoxic T lymphocyte and antibody responses in mice after intramuscular injection. , 1996, Journal of immunological methods.

[25]  H. Davis,et al.  DNA-mediated immunization to hepatitis B surface antigen: longevity of primary response and effect of boost. , 1996, Vaccine.

[26]  J. Reimann,et al.  Antibody and cytotoxic T-cell responses to soluble hepatitis B virus (HBV) S antigen in mice: implication for the pathogenesis of HBV-induced hepatitis , 1994, Journal of virology.

[27]  J. Reimann,et al.  Immunization with soluble hepatitis B virus surface protein elicits murine H-2 class I-restricted CD8+ cytotoxic T lymphocyte responses in vivo. , 1994, Journal of immunology.

[28]  J. Reimann,et al.  Selective stimulation of murine cytotoxic T cell and antibody responses by particulate or monomeric hepatitis B virus surface (S) antigen , 1994, European journal of immunology.

[29]  H. Davis,et al.  DNA-based immunization against the envelope proteins of the hepatitis B virus. , 1996, Journal of biotechnology.

[30]  H. Hauser,et al.  Dicistronic transcription units for gene expression in mammalian cells. , 1993, Gene.

[31]  M. Tao,et al.  Improvement of hepatitis B virus DNA vaccines by plasmids coexpressing hepatitis B surface antigen and interleukin-2 , 1997, Journal of virology.

[32]  M. Gossen,et al.  Temporal control of gene expression in transgenic mice by a tetracycline-responsive promoter. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[33]  F. Chisari.  Cytotoxic T cells and viral hepatitis. , 1997, The Journal of clinical investigation.

[34]  J. Reimann,et al.  Similar as well as distinct MHC class I‐binding peptides are generated by exogenous and endogenous processing of hepatitis B virus surface antigen , 1998, European journal of immunology.