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A novel DNA vaccine based on ubiquitin–proteasome pathway targeting ‘self’-antigens expressed in melanoma/melanocyte

Gene Therapy volume 12, pages 1049–1057 (2005)Cite this article

Abstract

Cancer vaccine that targets ‘self’-antigens expressed at high levels in tumor cells is a potentially useful immunotherapy, but immunological tolerance often defeats this strategy. Here, we describe the use of a naked DNA vaccine encoding a self tumor antigen, tyrosinase-related protein 2, to whose N-terminus ubiquitin is fused in a ‘nonremovable’ fashion. Unlike conventional DNA vaccines, this vaccine broke the tolerance and induced protective immunity to melanoma in C57BL/6 mice, as evaluated by tumor growth, survival rate and lung metastasis. The protective immunity was cancelled in the proteasome activator PA28α/β knockout mice. Moreover, this vaccination exhibited therapeutic effects on melanoma implanted before vaccination. Our findings provide evidence for the first time that naked DNA vaccines encoding a ubiquitin-fused self-antigen preferentially induce the main effector CD8+ T cells through efficient proteolysis mediated by the ubiquitin–proteasome pathway, and lead the way to strategies aimed at targeting tissue differentiation antigens expressed by tumors.

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References

  1. Rosenberg SA . Cancer vaccines based on the identification of genes encoding cancer regression antigens. Immunol Today 1997; 18: 175–182.

    Article  CAS  PubMed  Google Scholar 

  2. Wang RF et al. Identification of TRP-2 as a human tumor antigen recognized by cytotoxic T lymphocytes. J Exp Med 1996; 184: 2207–2216.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Weber LW et al. Tumor immunity and autoimmunity induced by immunization with homologous DNA. J Clin Invest 1998; 102: 1258–1264.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Overwijk WW et al. gp100/pmel 17 is a murine tumor rejection antigen: induction of ‘self’-reactive, tumoricidal T cells using high-affinity, altered peptide ligand. J Exp Med 1998; 188: 277–286.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Bowne WB et al. Coupling and uncoupling of tumor immunity and autoimmunity. J Exp Med 1999; 190: 1717–1722.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Tüting T et al. Induction of tumor antigen-specific immunity using plasmid DNA immunization in mice. Cancer Gene Ther 1999; 6: 73–80.

    Article  PubMed  Google Scholar 

  7. Steitz J et al. Genetic immunization with a melanocytic self-antigen linked to foreign helper sequences breaks tolerance and induces autoimmunity and tumor immunity. Gene Therapy 2002; 9: 208–213.

    Article  CAS  PubMed  Google Scholar 

  8. Engelhard VH et al. Antigen derived from melanocyte differentiation proteins: self-tolerance, autoimmunity, and use for cancer immunotherapy. Immunol Rev 2002; 188: 136–146.

    Article  CAS  PubMed  Google Scholar 

  9. Inaba K, Young JW, Steinman RM . Direct activation of CD8+ cytotoxic T lymphocytes by dendritic cells. J Exp Med 1987; 166: 182–194.

    Article  CAS  PubMed  Google Scholar 

  10. Sato M et al. Th1 cytokine-conditioned bone marrow-derived dendritic cells can bypass the requirement for Th functions during the generation of CD8+ CTL. J Immunol 2001; 167: 3687–3691.

    Article  CAS  PubMed  Google Scholar 

  11. Nishitani MA et al. A convenient cancer vaccine therapy with in vivo transfer of interleukin 12 expression plasmid using gene gun technology after priming with irradiated carcinoma cells. Cancer Gene Ther 2002; 9: 156–163.

    Article  CAS  PubMed  Google Scholar 

  12. Tanaka K, Kasahara M . The MHC class I ligand-generating system: roles of immunoproteasomes and the interferon-gamma-inducible proteasome activator PA28. Immunol Rev 1998; 163: 161–176.

    Article  CAS  PubMed  Google Scholar 

  13. Rock KL, York IA, Saric T, Goldberg AL . Protein degradation and the generation of MHC class I-presented peptides. In: Dixon FJ (ed). Advances in Immunology, Vol 80. Academic Press: Oxford, 2002, pp 1–71.

    Google Scholar 

  14. Kloetzel P-M . Antigen processing by the proteasome. Nat Rev Mol Cell Biol 2001; 2: 179–187.

    Article  CAS  PubMed  Google Scholar 

  15. Xiang R et al. An autologous oral DNA vaccine protects against murine melanoma. Proc Natl Acad Sci USA 2000; 97: 5492–5497.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Johnson ES, Ma PC, Ota IM, Varshavsky A . A proteolytic pathway that recognizes ubiquitin as a degradation signal. J Biol Chem 1995; 270: 17442–17456.

    Article  CAS  PubMed  Google Scholar 

  17. Zhang M et al. Ubiquitin-fusion degradation pathway plays an indispensable role in naked DNA vaccination with a chimeric gene encoding a syngeneic cytotoxic T lymphocyte epitope of melanocyte and green fluorescent protein. Immunology 2004; 112: 567–574.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Murata S et al. Immunoproteasome assembly and antigen presentation in mice lacking both PA28α and PA28β. EMBO J 2001; 20: 5898–5907.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. van den Eynde BJ, van der Bruggen P . T cell defined tumor antigens. Curr Opin Immunol 1997; 9: 684–693.

    Article  CAS  PubMed  Google Scholar 

  20. Rosenberg SA . A new era for cancer immunotherapy based on the genes that encode cancer antigens. Immunity 1999; 10: 281–287.

    Article  CAS  PubMed  Google Scholar 

  21. Bloom MB et al. Identification of tyrosinase-related protein 2 as a tumor rejection antigen for the B16 melanoma. J Exp Med 1997; 185: 453–459.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Lindauer M et al. The molecular basis of cancer immunotherapy by cytotoxic T lymphocytes. J Mol Med 1998; 76: 32–47.

    Article  CAS  PubMed  Google Scholar 

  23. Noppen C et al. Naturally processed and concealed HLA-A2.1-restricted epitopes from tumor-associated antigen tyrosinase-related protein-2. Int J Cancer 2000; 87: 241–246.

    Article  CAS  PubMed  Google Scholar 

  24. Rock KL et al. Inhibitors of the proteasome block the degradation of most cell proteins and the generation of peptides presented on MHC class I molecules. Cell 1994; 78: 761–771.

    Article  CAS  PubMed  Google Scholar 

  25. Cerundolo V et al. The proteasome-specific inhibitor lactocystin blocks presentation of cytotoxic T lymphocyte epitopes in human and murine cells. Eur J Immunol 1997; 27: 336–341.

    Article  CAS  PubMed  Google Scholar 

  26. Craiu A et al. Lactacystin and clasto-lactacystin b-lactone modify multiple proteasome β-subunits and inhibit intracellular protein degradation and major histocompatibility complex class I antigen presentation. J Biol Chem 1997; 272: 13437–13445.

    Article  CAS  PubMed  Google Scholar 

  27. Hershko A, Ciechanover A . The ubiquitin system. Annu Rev Biochem 1998; 67: 425–479.

    Article  CAS  PubMed  Google Scholar 

  28. Rodriguez F, Zhang J, Whitton JL . DNA immunization: ubiquitination of a viral protein enhances cytotoxic T-lymphocyte induction and antiviral protection but abrogates antibody induction. J Virol 1997; 71: 8497–8503.

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Leitner WW et al. Alphavirus-based DNA vaccine breaks immunological tolerance by activating innate antiviral pathway. Nat Med 2003; 9: 33–39.

    Article  CAS  PubMed  Google Scholar 

  30. Overwijk WW et al. Vaccination with a recombinant vaccinia virus encoding a ‘self’ antigen induces autoimmune vitiligo and tumor cell destruction in mice: requirement for CD4+ T lymphocytes. Proc Natl Acad Sci USA 1999; 96: 2982–2987.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Mackensen A et al. Phase I study in melanoma patients of a vaccine with peptide-pulsed dendritic cells generated in vitro from CD34+ hematopoietic progenitor cells. Int J Cancer 2000; 86: 385–392.

    Article  CAS  PubMed  Google Scholar 

  32. Bronte V et al. Genetic vaccination with ‘self’ tyrosinase-related protein 2 causes melanoma eradication but not vitiligo. Cancer Res 2000; 60: 253–258.

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Sakai T et al. Gene gun-mediated delivery of an interleukin-12 expression plasmid protects against infections with the intracellular protozoan parasites Leishmaniamajor and Trypanosoma cruzi in mice. Immunology 2000; 99: 615–624.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Sakai T et al. Gene gun-based co-immunization of merozoit surface protein-1 cDNA with IL-12 expression plasmid confers protection against lethal Plasmodium yoelii in A/J mice. Vaccine 2003; 21: 1432–1444.

    Article  CAS  PubMed  Google Scholar 

  35. Nanni P et al. Interleukin 12 gene therapy of MHC-negative murine melanoma metastases. Cancer Res 1998; 58: 1225–1230.

    CAS  PubMed  Google Scholar 

  36. Matzinger P . The JAM test. A simple assay for DNA fragmentation and cell death. J Immunol Methods 1991; 145: 185–192.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by grants-in-aid from the Ministry of Education, Culture, Sport, Science, and Technology of Japan (15019075, 15025255, 15390136, 15659265).

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  1. M Zhang and C Obata: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Microbiology and Immunology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan

    M Zhang, H Hisaeda, K Ishii, Y Li, B Chou, T Imai, X Duan & K Himeno

  2. Department of Dermatology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan

    C Obata & M Furue

  3. Department of Molecular Oncology, The Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan

    S Murata, T Chiba & K Tanaka

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Zhang, M., Obata, C., Hisaeda, H. et al. A novel DNA vaccine based on ubiquitin–proteasome pathway targeting ‘self’-antigens expressed in melanoma/melanocyte. Gene Ther 12, 1049–1057 (2005). https://doi.org/10.1038/sj.gt.3302490

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