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.

  • Research Article
  • Published:

Flt-3 ligand as adjuvant for DNA vaccination augments immune responses but does not skew TH1/TH2 polarization

Gene Therapy volume 11, pages 1048–1056 (2004)Cite this article

Abstract

Since transfection of dendritic cells (DC) plays a key role in DNA vaccination, in vivo expansion of DC might be a tool to increase vaccine efficacy. We asked whether Fms-like tyrosine kinase-3 ligand (Flt-3L), a growth factor for DC, can be used as an adjuvant for DNA vaccination. Beta-galactosidase (β-gal) was used as a model antigen in C57BL/6 mice. Mice were immunized i.m. with DNA coding for β-gal with or without additional injection of Flt-3L. In both cases, antigen-specific CD4+ and CD8+ T cells were detectable after vaccination. Compared with DNA alone, additional administration of Flt-3L led to a significant increase in the antigen-specific proliferative response. However, increased cytotoxicity by T cells was not observed. The cytokines secreted by splenocytes of immunized mice upon in vitro stimulation with antigen had a TH2 profile. Humoral responses against β-gal preferentially consisted of IgG1 antibodies. Analysis of DC from Flt-3L-treated mice revealed an immature phenotype with low or absent expression levels of CD80, CD86 and CD40. We conclude that Flt-3L does not generally skew immune responses towards a TH1 type. More likely, factors determined by the antigen and/or the vaccination procedure itself are crucial for the resulting type of immune response. Flt-3L – under circumstances such as the one we have investigated – can also lead to suppression of TH1 T cell immunity, possibly by expansion of immature/unactivated DC.

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
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9

Similar content being viewed by others

References

  1. Condon C et al. DNA-based immunization by in vivo transfection of dendritic cells. Nat Med 1996; 2: 1122–1128.

    Article  CAS  Google Scholar 

  2. Gurunathan S, Klinman DM, Seder RA . DNA Vaccines: immunology, application and optimization. Ann Rev Immunol 2000; 18: 927–974.

    Article  CAS  Google Scholar 

  3. Scheerlinck JY . Genetic adjuvants for DNA vaccines. Vaccine 2001; 19: 2647–2656.

    Article  CAS  Google Scholar 

  4. Gilliland DG, Griffin JD . The roles of FLT3 in hematopoiesis and leukemia. Blood 2002; 100: 1532–1542.

    Article  CAS  Google Scholar 

  5. Maraskovsky E et al. Dramatic increase in the numbers of functionally mature dendritic cells in Flt3 ligand-treated mice: multiple dendritic cell subpopulations identified. J Exp Med 1996; 184: 1953–1962.

    Article  CAS  Google Scholar 

  6. Maraskovsky E et al. In vivo generation of human dendritic cell subsets by Flt3 ligand. Blood 2000; 96: 878–884.

    CAS  Google Scholar 

  7. Oughton JA, Kerkvliet NL . Novel phenotype associated with in vivo activated CTL. Clin Immunol 1999; 90: 323–333.

    Article  CAS  Google Scholar 

  8. Inaba K et al. Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor. J Exp Med 1992; 176: 1693–1702.

    Article  CAS  Google Scholar 

  9. Schuler G, Schuler-Thurner B, Steinman RM . The use of dendritic cells in cancer immunotherapy. Curr Opin Immunol 2003; 15: 138–147.

    Article  CAS  Google Scholar 

  10. Doe B et al. Induction of cytotoxic T lymphocytes by intramuscular immunization with plasmid DNA is facilitated by bone marrow-derived cells. Proc Natl Acad Sci USA 1996; 57: 873–883.

    Google Scholar 

  11. Casares S et al. Antigen presentation by dendritic cells after immunization with DNA encoding a major histocompatibility complex class II-restricted viral epitope. J Exp Med 1997; 186: 1481–1486.

    Article  CAS  Google Scholar 

  12. Porgador A et al. Predominant role for directly transfected dendritic cells in antigen presentation to CD8+ T cells after gene gun immunization. J Exp Med 1998; 188: 1075–1082.

    Article  CAS  Google Scholar 

  13. Zöller M, Christ O . Prophylactic tumor vaccination: comparison of effector mechanisms initiated by protein versus DNA vaccination. J Immunol 2001; 166: 3440–3450.

    Article  Google Scholar 

  14. Sun X et al. Co-expression of granulocyte–macrophage colony-stimulating factor with antigen enhances humoral and tumor immunity after DNA vaccination. Vaccine 2002; 20: 1466–1474.

    Article  CAS  Google Scholar 

  15. Parajuli P et al. Immunization with wild-type p53 gene sequences coadministered with Flt3 ligand induces an antigen-specific type 1 T-cell response. Cancer Res 2001; 61: 8227–8234.

    CAS  PubMed  Google Scholar 

  16. Moser M . Dendritic cells in immunity and tolerance – do they display opposite functions? Immunity 2003; 19: 5–8.

    Article  CAS  Google Scholar 

  17. Lynch DH et al. Flt3 ligand induces tumor regression and antitumor immune responses in vivo. Nat Med 1997; 3: 625–631.

    Article  CAS  Google Scholar 

  18. Pawlowska AB et al. In vitro tumor-pulsed or in vivo Flt3 ligand-generated dendritic cells provide protection against acute myelogenous leukemia in nontransplanted or syngeneic bone marrow-transplanted mice. Blood 2001; 97: 1474–1482.

    Article  CAS  Google Scholar 

  19. Braun SE et al. Flt3 ligand antitumor activity in a murine breast cancer model: a comparison with granulocyte–macrophage colony-stimulating factor and a potential mechanism of action. Hum Gene Ther 1999; 10: 2141–2151.

    Article  CAS  Google Scholar 

  20. Hung CF et al. Enhancement of DNA vaccine potency by linkage of antigen gene to a gene encoding the extracellular domain of fms-like tyrosine kinase 3-ligand. Cancer Res 2001; 61: 1080–1088.

    CAS  PubMed  Google Scholar 

  21. Fong CL, Hui KM . Generation of potent and specific cellular immune responses via in vivo stimulation of dendritic cells by pNGVL3-hFLex plasmid DNA and immunogenic peptides. Gene Therapy 2002; 9: 1127–1138.

    Article  CAS  Google Scholar 

  22. Liu Y et al. Dendritic cells engineered to express the Flt3 ligand stimulate type I immune response, and induce enhanced cytotoxic T and natural killer cell cytotoxicities and antitumor immunity. J Gene Med 2003; 5: 668–680.

    Article  CAS  Google Scholar 

  23. Smith JR, Thackray AM, Bujdoso R . Reduced herpes simplex virus type 1 latency in Flt-3 ligand-treated mice is associated with enhanced numbers of natural killer and dendritic cells. Immunology 2001; 102: 352–358.

    Article  CAS  Google Scholar 

  24. Pulendran B et al. Prevention of peripheral tolerance by a dendritic cell growth factor: flt3 ligand as an adjuvant. J Exp Med 1998; 188: 2075–2082.

    Article  CAS  Google Scholar 

  25. Kim EM, Sivanandham M, Stavropoulos CI, Wallack MK . Adjuvant effect of a Flt3 ligand (FL) gene-transduced xenogeneic cell line in a Murine Colon Cancer Model. J Surg Res 2002; 108: 148–156.

    Article  CAS  Google Scholar 

  26. Kwon TK, Park JW . Intramuscular co-injection of naked DNA encoding HBV core antigen and Flt3 ligand suppresses anti-HBc antibody response. Immunol Lett 2002; 81: 229–234.

    Article  CAS  Google Scholar 

  27. Evans TG, Hasan M, Galibert L, Caron D . The use of Flt3 ligand as an adjuvant for hepatitis B vaccination of healthy adults. Vaccine 2002; 21: 322–329.

    Article  CAS  Google Scholar 

  28. McNeel DG et al. Pilot study of an HLA-A2 peptide vaccine using flt3 ligand as a systemic vaccine adjuvant. J Clin Immunol 2003; 23: 62–72.

    Article  CAS  Google Scholar 

  29. Viney JL et al. Expanding dendritic cells in vivo enhances the induction of oral tolerance. J Immunol 1998; 160: 5815–5825.

    CAS  PubMed  Google Scholar 

  30. Yunusov MY et al. FLT3 ligand promotes engraftment of allogeneic hematopoietic stem cells without significant graft-versus-host disease. Transplantation 2003; 75: 933–940.

    Article  CAS  Google Scholar 

  31. Ciavarra RP et al. Impact of the tumor microenvironment on host infiltrating cells and the efficacy of Flt3-ligand combination immunotherapy evaluated in a treatment model of mouse prostate cancer. Cancer Immunol Immunother 2003; 52: 535–545.

    Article  CAS  Google Scholar 

  32. Langenkamp A et al. Kinetics of dendritic cell activation: impact on priming of TH1, TH2 and nonpolarized cells. Nat Immunol 2000; 1: 311–316.

    Article  CAS  Google Scholar 

  33. Boonstra A et al. Flexibility of mouse classical and plasmacytoid-derived dendritic cells in directing T helper type 1 and 2 cell development: dependency on antigen dose and differential toll-like receptor ligation. J Exp Med 2003; 197: 101–109.

    Article  CAS  Google Scholar 

  34. George TC, Bilsborough J, Viney JL, Norment AM . High antigen dose and activated dendritic cells enable Th cells to escape regulatory T cell-mediated suppression in vitro. Eur J Immunol 2003; 33: 502–511.

    Article  CAS  Google Scholar 

  35. Robinson HL . DNA vaccines: basic mechanisms and immune response. Int J Mol Med 1999; 4: 549–555.

    CAS  PubMed  Google Scholar 

  36. Shivanthi P et al. The ability of murine dendritic cell subsets to direct T helper cell differentiation is dependent on microbial signals. Eur J Immunol 2003; 33: 101–107.

    Article  Google Scholar 

  37. Merad M, Sugie T, Engleman EG, Fong L . In vivo manipulation of dendritic cells to induce therapeutic immunity. Blood 2002; 99: 1676–1682.

    Article  CAS  Google Scholar 

  38. Sato M, Iwakabe K, Kimura S, Nishimura T . Functional skewing of bone marrow-derived dendritic cells by Th1- or Th2-inducing cytokines. Immunol Lett 1999; 67: 63–68.

    Article  CAS  Google Scholar 

  39. Mosca PJ et al. Multiple signals are required for maturation of human dendritic cells mobilized in vivo with Flt3 ligand. J Leukocyte Biol 2002; 72: 546–553.

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Hematology, Oncology and Tumorimmunology, Charité – University Medicine Berlin, Campus Berlin-Buch, Berlin, Germany

    J Westermann, T Nguyen-Hoai, A Mollweide, G Richter, O Schmetzer, B Dörken & A Pezzutto

  2. Max Delbrück Center for Molecular Medicine, Berlin, Germany

    J Westermann, T Nguyen-Hoai, A Mollweide, G Richter, O Schmetzer, H-J Kim, Th Blankenstein, B Dörken & A Pezzutto

  3. Institute of Immunology, Charité – University Medicine Berlin, Campus Benjamin Franklin, Berlin, Germany

    Th Blankenstein

Authors
  1. J Westermann
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T Nguyen-Hoai
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A Mollweide
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G Richter
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. O Schmetzer
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. H-J Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Th Blankenstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. B Dörken
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. A Pezzutto
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Westermann, J., Nguyen-Hoai, T., Mollweide, A. et al. Flt-3 ligand as adjuvant for DNA vaccination augments immune responses but does not skew TH1/TH2 polarization. Gene Ther 11, 1048–1056 (2004). https://doi.org/10.1038/sj.gt.3302261

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3302261

Keywords

This article is cited by

Search

Advanced search

Quick links