Abstract
In DNA vaccination, an exciting new immunization technique with potential applications in clinical medicine, expression plasmid DNA containing antigen-encoding sequences cloned under heterologous promoter control are delivered by techniques that lead in vivo to antigen expression in transfected cells. DNA vaccination efficiently primes both humoral and cellular immune responses. We developed a novel expression system for DNA vaccines in which a fusion protein with a small, N-terminal, viral DnaJ-like sequence (J domain) is translated in frame with C-terminal antigen-encoding sequences. The J domain stable bind to constitutively expressed, cytosolic stress protein hsp73 and triggers intracellular accumulation of antigen/hsp73 complexes. The system supports enhanced expression of chimeric antigens of >800 residues in length in immunogenic form. A unique advantage of the system is that even unstable or toxic proteins (or protein domains) can be expressed. We describe the design of DNA vaccines expressing antigens with a stress protein-capturing domain and characterize the immunogenicity of the antigens produced by this expression system.
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References
Reimann, J. and Schirmbeck, R. (2004) DNA vaccines expressing antigens with a stress protein-capturing domain display enhanced immunogenicity. Immunol. Rev. 199, 54–67.
Echeverria, P., Dran, G., Pereda, G., et al. (2001) Analysis of the adjuvant effect of recombinant Leishmania infantum Hsp83 protein as a tool for vaccination. Immunol. Lett. 76, 107–110.
Wang, Y., Kelly, C. G., Karttunen, J. T., et al. (2001) CD40 is a cellular receptor mediating mycobacterial heat shock protein 70 stimulation of CC-chemokines. Immunity 15, 971–983.
Bethke, K., Staib, F., Distler, M., et al. (2002) Different efficiency of heat shock proteins (HSP) to activate human monocytes and dendritic cells: superiority of HSP60. J. Immunol. 169, 6141–6148.
Panjwani, N. N., Popova, L., and Srivastava, P. (2002) Heat shock proteins gp96 and hsp70 activate the release of nitric oxide by APCs. J. Immunol. 168, 2997–3003.
Millar, D. G., Garza, K. M., Odermatt, B., et al. (2003) Hsp70 promotes antigen-presenting cell function and converts T-cell tolerance to autoimmunity in vivo. Nat. Med. 9, 1469–1476.
Palliser, D., Huang, Q., Hacohen, N., et al. (2004) A role for toll-like receptor 4 in dendritic cell activation and cytolytic CD8+ T cell differentiation in response to a recombinant heat shock fusion protein. J. Immunol. 172, 2885–2893.
Suzue, K., Zhou, X., Eisen, H. N., and Young, R. (1997) Heat shock fusion proteins as vehicles for antigen delivery into the major histocompatibility complex class I presentation pathway. Proc. Natl. Acad. Sci. USA 94, 13,146–13,151.
Cho, B. K., Palliser, D., Guillen, E., et al. (2000) A proposed mechanism for the induction of cytotoxic T lymphocyte production by heat shock fusion proteins. Immunity 12, 263–272.
Huang, Q., Richmond, J. F., Suzue, K., Eisen, H. N., and Young, R. A. (2000) In vivo cytotoxic T lymphocyte elicitation by mycobacterial heat shock protein 70 fusion proteins maps to a discrete domain and is CD4+ T cell independent. J. Exp. Med. 191, 403–408.
Arnold-Schild, D., Kleist, C., Welschof, M., et al. (2000) One-step single-chain Fv recombinant antibody-based purification of gp96 for vaccine development. Cancer Res. 60, 4175–4178.
Srivastava, P. (2000) Immunotherapy of human cancer: lessons from mice. Nat. Immunol. 1, 363–366.
Srivastava, P. and Jaikaria, N. S. (2001) Methods of purification of heat shock protein-peptide complexes for use as vaccines against cancers and infectious diseases. Methods Mol. Biol. 156, 175–186.
Schirmbeck, R., Böhm, W., and Reimann, J. (1997) Stress protein (hsp73)-mediated, TAP-independent processing of endogenous, truncated SV40 large T antigen for Db-restricted peptide presentation. Eur. J. Immunol. 27, 2016–2023.
Schirmbeck, R., Kwissa, M., Fissolo, N., Elkholy, S., Riedl, P., and Reimann, J. (2002) Priming polyvalent immunity by DNA vaccines expressing chimeric antigens with a stress protein-capturing, viral J-domain. FASEB J. 16, 1108–1110.
Schirmbeck, R. and Reimann, J. (1994) Peptide transporter-independent, stress protein-mediated endosomal processing of endogenous protein antigens for major histocompatibility complex class I presentation. Eur. J. Immunol. 24, 1478–1486.
Schirmbeck, R., Stober, D., El Kholy, S., Riedl, P., and Reimann, J. (2002) The immunodominant, Ld-restricted T cell response to hepatitis B surface antigen (HBsAg) efficiently suppresses T cell priming to multiple Dd-, Kd-, and Kb-restricted HBsAg epitopes. J. Immunol. 168, 6253–6262.
Corr, M., Lee, D. J., Carson, D. A., and Tighe, H. (1996) Gene vaccination with naked plasmid DNA: mechanism of CTL priming. J. Exp. Med. 184, 1555–1560.
Doe, B., Selby, M., Barnett, S., Baenziger, J., and Walker, C. M. (1996) Induction of cytotoxic T lymphocytes by intramuscular immunization with plasmid DNA is facilitated by bone marrow-derived cells. Proc. Natl. Acad. Sci. USA 93, 8578–8583.
Ulmer, J. B., Deck, R. R., DeWitt, C. M., Donnhly, J. I., and Liu, M. A. (1996) Generation of MHC class I-restricted cytotoxic T lymphocytes by expression of a viral protein in muscle cells: antigen presentation by non-muscle cells. Immunology 89, 59–67.
Fu, T. M., Ulmer, J. B., Caulfield, M. J., et al. (1997) Priming of cytotoxic T lymphocytes by DNA vaccines: requirement for professional antigen presenting cells and evidence for antigen transfer from myocytes. Mol. Med. 3, 362–371.
Iwasaki, A., Torres, C. A., Ohashi, P. S., Robinson, H. L., and Barber, B. H. (1997) The dominant role of bone marrow-derived cells in CTL induction following plasmid DNA immunization at different sites. J. Immunol. 159, 11–14.
Corr, M., von-Damm, A., Lee, D. J., and Tighe, H. (1999) In vivo priming by DNA injection occurs predominantly by antigen transfer. J. Immunol. 163, 4721–4727.
Kammerer, R., Stober, D., Riedl, P., Oehninger, C., Schirmbeck, R., and Reimann, J. (2002) Noncovalent association with stress protein facilitates cross-priming of CD8+ T cells to tumor cell antigens by dendritic cells. J. Immunol. 168, 108–117.
Schirmbeck, R., Gerstner, O., and Reimann, J. (1999) Truncated or chimeric endogenous protein antigens gain immunogenicity for B cells by stress protein-facilitated expression. Eur. J. Immunol. 29, 1740–1749.
El Kholy, S., Riedl, P., Kwissa, M., Reimann, J., and Schirmbeck, R. (2002) Selective expression of immunogenic, VLP-derived epitopes binding antibodies. Intervirology 45, 251–259.
Riedl, P., El Kholy, S., Reimann, J., and Schirmbeck, R. (2002) Priming biologically active antibody responses against an isolated, conformational viral epitope by DNA vaccination. J. Immunol. 169, 1251–1260.
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Riedl, P., Fissolo, N., Reimann, J., Schirmbeck, R. (2006). A Stress Protein-Facilitated Antigen Expression System for Plasmid DNA Vaccines. In: Saltzman, W.M., Shen, H., Brandsma, J.L. (eds) DNA Vaccines. Methods in Molecular Medicine™, vol 127. Humana Press. https://doi.org/10.1385/1-59745-168-1:41
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DOI: https://doi.org/10.1385/1-59745-168-1:41
Publisher Name: Humana Press
Print ISBN: 978-1-58829-484-5
Online ISBN: 978-1-59745-168-0
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