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.

  • Full Paper
  • Published:

Effect on antibody and T-cell responses of mixing five GMP-produced DNA plasmids and administration with plasmid expressing GM-CSF

Genes & Immunity volume 5, pages 553–561 (2004)Cite this article

Abstract

One potential benefit of DNA vaccines is the capacity to elicit antibody and T-cell responses against multiple antigens at the same time by mixing plasmids expressing different proteins. A possible negative effect of such mixing is interference among plasmids regarding immunogenicity. In preparation for a clinical trial, we assessed the immunogenicity of GMP-produced plasmids encoding five Plasmodium falciparum proteins, PfCSP, PfSSP2, PfEXP1, PfLSA1, and PfLSA3, given as a mixture, or alone. The mixture induced higher levels of antibodies against whole parasites than did the individual plasmids, but was associated with a decrease in antibodies to individual P. falciparum proteins. T-cell responses were in general decreased by administration of the mixture. Immune responses to individual plasmids and mixtures were generally higher in inbred mice than in outbreds. In inbred BALB/c and C57BL/6 mice, coadministration of a plasmid expressing murine granulocyte–macrophage colony-stimulating factor (mGM-CSF), increased antibody and T-cell responses, but in outbred CD-1 mice, coadministration of mGM-CSF was associated with a decrease in antibody responses. Such variability in data from studies in different strains of mice underscores the importance of genetic background on immune response and carefully considering the goals of any preclinical studies of vaccine mixtures planned for human trials.

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

Similar content being viewed by others

References

  1. Hoffman SL, Franke ED, Hollingdale MR, Druilhe P . Attacking the infected hepatocyte. In: Hoffman SL (ed). Malaria Vaccine Development: A Multi-Immune Response Approach. ASM Press: Washington, DC, 1996, pp 35–75.

    Google Scholar 

  2. Sedegah M, Hedstrom R, Hobart P, Hoffman SL . Protection against malaria by immunization with plasmid DNA encoding circumsporozoite protein. Proc Natl Acad Sci USA 1994; 91: 9866–9870.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Doolan DL, Sedegah M, Hedstrom RC, Hobart P, Charoenvit Y, Hoffman SL . Circumventing genetic restriction of protection against malaria with multi-gene DNA immunization: CD8+ T cell, interferon-gamma, and nitric oxide dependent immunity. J Exp Med 1996; 183: 1739–1746.

    Article  CAS  PubMed  Google Scholar 

  4. Hedstrom RC, Doolan DL, Wang R et al. In vitro expression and in vivo immunogenicity of Plasmodium falciparum pre-erythrocytic stage DNA vaccines. Int J Mol Med 1998; 2: 29–38.

    CAS  PubMed  Google Scholar 

  5. Aguiar JC, Hedstrom RC, Rogers WO et al. Enhancement of the immune response in rabbits to a malaria DNA vaccine by immunization with a needle-free jet device. Vaccine 2001; 20: 275–280.

    Article  CAS  PubMed  Google Scholar 

  6. Jones TR, Narum DL, Gozalo AS et al. Protection of Aotus monkeys by Plasmodium falciparum EBA-175 prime-protein boost immunization regimen. J Infect Dis 2001; 183: 303–312.

    Article  CAS  PubMed  Google Scholar 

  7. Jones TR, Gramzinski RA, Aguiar JC et al. Absence of antigenic competition in Aotus monkeys immunized with Plasmodium falciparum DNA vaccines delivered as a mixture. Vaccine 2002; 20: 1675–1680.

    Article  CAS  PubMed  Google Scholar 

  8. Rogers WO, Baird JK, Kumar A et al. Multistage multiantigen heterologous prime boost vaccine for Plasmodium knowlesi malaria provides partial protection in rhesus macaques. Infect Immun 2001; 69: 5565–5572.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Wang R, Doolan DL, Charoenvit Y et al. Simultaneous induction of multiple antigen-specific cytotoxic T lymphocytes in nonhuman primates by immunization with a mixture of four Plasmodium falciparum DNA plasmids. Infect Immun 1998; 66: 4193–4202.

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Wang R, Doolan DL, Le TP et al. Induction of antigen-specific cytotoxic T lymphocytes in humans by a malaria DNA vaccine. Science 1998; 282: 476–480.

    Article  CAS  PubMed  Google Scholar 

  11. Le TP, Coonan KM, Hedstrom RC et al. Safety, tolerability and humoral immune responses after intramuscular administration of a malaria DNA vaccine to healthy adult volunteers. Vaccine 2000; 18: 1893–1901.

    Article  CAS  PubMed  Google Scholar 

  12. Wang R, Epstein J, Baraceros FM et al. Induction of CD4(+) T cell-dependent CD8(+) type 1 responses in humans by a malaria DNA vaccine. Proc Natl Acad Sci USA 2001; 98: 10817–10822.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Weiss WR, Ishii KJ, Hedstrom RC et al. A plasmid encoding murine granulocyte–macrophage colony-stimulating factor increases protection conferred by a malaria DNA vaccine. J Immunol 1998; 161: 2325–2332.

    CAS  PubMed  Google Scholar 

  14. Sedegah M, Weiss W, Sacci Jr JB et al. Improving protective immunity induced by DNA-based immunization: priming with antigen and GM-CSF encoding plasmid DNA and boosting with antigen expressing recombinant poxvirus. J Immunol 2000; 164: 5905–5912.

    Article  CAS  PubMed  Google Scholar 

  15. Kumar S, Epstein JE, Richie TL et al. A multilateral effort to develop DNA vaccines against falciparum malaria. Trends Parasitol 2002; 18: 129–135.

    Article  CAS  PubMed  Google Scholar 

  16. Hoffman SL, Doolan DL . Malaria vaccines-targeting infected hepatocytes. Nat Med 2000; 6: 1218–1219.

    Article  CAS  PubMed  Google Scholar 

  17. Malik A, Egan JE, Houghten RA, Sadoff JC, Hoffman SL . Human cytotoxic T lymphocytes against the Plasmodium falciparum circumsporozoite protein. Proc Natl Acad Sci USA 1991; 88: 3300–3304.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Sedegah M, Sim BKL, Mason C et al. Naturally acquired CD8+ cytotoxic T lymphocytes against the Plasmodium falciparum circumsporozoite protein. J Immunol 1992; 149: 966–971.

    CAS  PubMed  Google Scholar 

  19. Weiss WR, Berzovsky JA, Houghten RA, Sedegah M, Hollingdale M, Hoffman SL . A T cell clone directed at the circumsporozoite protein which protects mice against both Plasmodium yoelii and Plasmodium berghei. J Immunol 1992; 149: 2103–2109.

    CAS  PubMed  Google Scholar 

  20. Doolan DL, Hoffman SL . The complexity of protective immunity against liver-stage malaria. J Immunol 2000; 165: 1453–1462.

    Article  CAS  PubMed  Google Scholar 

  21. Perlaza BL, Sauzet JP, Balde AT et al. Long synthetic peptides encompassing the Plasmodium falciparum LSA3 are the target of human B and T cells and are potent inducers of B helper, T helper and cytolytic T cell responses in mice. Eur J Immunol 2001; 31: 2200–2209.

    Article  CAS  PubMed  Google Scholar 

  22. Meraldi V, Nebie I, Moret R et al. Recognition of synthetic polypeptides corresponding to the N- and C-terminal fragments of Plasmodium falciparum Exp-1 by T-cells and plasma from human donors from African endemic areas. Parasite Immunol 2002; 24: 141–150.

    Article  PubMed  Google Scholar 

  23. Sedegah M, Charoenvit Y, Minh L et al. Reduced immunogenicity of DNA vaccine plasmids in mixtures. Gene Therapy 2004; 11: 448–456.

    Article  CAS  PubMed  Google Scholar 

  24. Kumar S, Villinger F, Oakley M et al. A DNA vaccine encoding the 42 kDa C-terminus of merozoite surface protein 1 of Plasmodium falciparum induces antibody, interferon-gamma and cytotoxic T cell responses in rhesus monkeys: immuno-stimulatory effects of granulocyte macrophage-colony stimulating factor. Immunol Lett 2002; 81: 13–24.

    Article  CAS  PubMed  Google Scholar 

  25. Kolodny N, Kitov S, Vassell MA et al. Two-step chromatographic purification of recombinant Plasmodium falciparum circumsporozoite protein from Escherichia coli. J Chromatogr B 2001; 762: 77–86.

    Article  CAS  Google Scholar 

  26. Daubersies P, Thomas AW, Millet P et al. Protection against Plasmodium falciparum malaria in chimpanzees by immunization with the conserved pre-erythrocytic liver-stage antigen 3. Nat Med 2000; 6: 1258–1263.

    Article  CAS  PubMed  Google Scholar 

  27. Caspers P, Etlinger H, Matile H, Pink JR, Stuber D, Takacs B . A Plasmodium falciparum malaria vaccine candidate which contains epitopes from the circumsporozoite protein and a blood stage antigen, 5.1. Mol Biochem Parasitol 1991; 47: 143–150.

    Article  CAS  PubMed  Google Scholar 

  28. Charoenvit Y, Mellouk S, Cole C et al. Monoclonal, but not polyclonal antibodies protect against Plasmodium yoeli sporozoites. J Immunol 1991; 146: 1020–1025.

    CAS  PubMed  Google Scholar 

  29. Blum Tirouvanziam U, Servis C, Habluetzel A et al. Localization of HLA-A2.1-restricted T cell epitopes in the circumsporozoite protein of Plasmodium falciparum. J Immunol 1995; 154: 3922–3931.

    CAS  PubMed  Google Scholar 

  30. Wizel B, Rogers WO, Houghten RA, Lanar DE, Tine JA, Hoffman SL . Induction of murine cytotoxic T lymphocytes against Plasmodium falciparum sporozoite surface protein 2. Eur J Immunol 1994; 24: 1487–1495.

    Article  CAS  PubMed  Google Scholar 

  31. Miyahira Y, Murata K, Rodriguez D et al. Quantification of antigen specific CD8+ T cells using an ELISPOT assay. J Immunol Methods 1995; 181: 45–54.

    Article  CAS  PubMed  Google Scholar 

  32. Sedegah M, Jones TR, Kaur M et al. Boosting with recombinant vaccinia increases immunogenicity and protective efficacy of malaria DNA vaccine. Proc Natl Acad Sci USA 1998; 95: 7648–7653.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We are grateful to Romeo Wallace, Victoria Fallame Majam, and Harini Ganeshan for technical support. We thank Dr WR Weiss for discussions and editorial assistance. The experiments reported here were carried out according to the principles set forth in the ‘Guide for the Care and Use of Laboratory Animals’, Institute of Laboratory Animals Resources, National Research Council, National Academy Press, 1996. This study was supported by Naval Medical Research Center work unit #A2284. The opinions and assertions herein are the private ones of the authors and are not to be construed as official or as reflecting the views of the US Navy, the US Department of Defense, or the US Government.

Author information

Author notes

  1. J Sacci

    Present address: Department of Microbiology, The University of Maryland School of Medicine, Baltimore, MD, USA

  2. R Hedstrom

    Present address: DynPort Vaccine Company LLC, Frederick, MD, USA

  3. S Kumar

    Present address: CBER, Food and Drug Administration, Rockville, MD, USA

  4. SL Hoffman

    Present address: Sanaria Inc., 12111 Parklawn Drive, Rockville, MD, 20852, USA

Authors and Affiliations

  1. Malaria Program, Naval Medical Research Center, Silver Spring, MD, USA

    M Sedegah, Y Charoenvit, J Aguiar, J Sacci, R Hedstrom, S Kumar, A Belmonte, TR Jones, E Abot, JE Epstein, TL Richie, DJ Carucci & SL Hoffman

  2. Walter Reed Army Institute of Research, Silver Spring, MD, USA

    DE Lanar

  3. Parasitologie Biomedicale, Institut Pasteur, France

    P Druilhe

  4. Institute of Biochemistry, University of Lausanne, Epalinges, Switzerland

    G Corradin

Authors
  1. M Sedegah
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Y Charoenvit
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J Aguiar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J Sacci
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. R Hedstrom
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. S Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. A Belmonte
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. DE Lanar
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. TR Jones
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. E Abot
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. P Druilhe
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. G Corradin
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. JE Epstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. TL Richie
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. DJ Carucci
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. SL Hoffman
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to SL Hoffman.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sedegah, M., Charoenvit, Y., Aguiar, J. et al. Effect on antibody and T-cell responses of mixing five GMP-produced DNA plasmids and administration with plasmid expressing GM-CSF. Genes Immun 5, 553–561 (2004). https://doi.org/10.1038/sj.gene.6364125

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gene.6364125

Keywords

This article is cited by

Search

Advanced search

Quick links