Cancer Immunology, Immunotherapy

, Volume 60, Issue 3, pp 327–337 | Cite as

A long peptide from MELOE-1 contains multiple HLA class II T cell epitopes in addition to the HLA-A*0201 epitope: an attractive candidate for melanoma vaccination

  • Anne Rogel
  • Virginie Vignard
  • Mathilde Bobinet
  • Nathalie Labarriere
  • François Lang
Original Article


CD4+ T cells contribute importantly to the antitumor T cell response, and thus, long peptides comprising CD4 and CD8 epitopes may be efficient cancer vaccines. We have previously identified an overexpressed antigen in melanoma, MELOE-1, presenting a CD8+ T cell epitope, MELOE-136–44, in the HLA-A*0201 context. A T cell repertoire against this epitope is present in HLA-A*0201+ healthy subjects and melanoma patients and the adjuvant injection of TIL containing MELOE-1 specific CD8+ T cells to melanoma patients was shown to be beneficial. In this study, we looked for CD4+ T cell epitopes in the vicinity of the HLA-A*0201 epitope. Stimulation of PBMC from healthy subjects with MELOE-126–46 revealed CD4 responses in multiple HLA contexts and by cloning responsive CD4+ T cells, we identified one HLA-DRβ1*1101-restricted and one HLA-DQβ1*0603-restricted epitope. We showed that the two epitopes could be efficiently presented to CD4+ T cells by MELOE-1-loaded dendritic cells but not by MELOE-1+ melanoma cell-lines. Finally, we showed that the long peptide MELOE-122–46, containing the two optimal class II epitopes and the HLA-A*0201 epitope, was efficiently processed by DC to stimulate CD4+ and CD8+ T cell responses in vitro, making it a potential candidate for melanoma vaccination.


T cell epitope CD4+ T cells Melanoma Vaccination 

Supplementary material

262_2010_938_MOESM1_ESM.pdf (262 kb)
Supplementary material 1 (PDF 261 kb)


  1. 1.
    Rosenberg SA, Yang JC, Restifo NP (2004) Cancer immunotherapy: moving beyond current vaccines. Nat Med 10:909–915CrossRefPubMedGoogle Scholar
  2. 2.
    Melief CJ, van der Burg SH (2008) Immunotherapy of established (pre)malignant disease by synthetic long peptide vaccines. Nat Rev Cancer 8:351–360CrossRefPubMedGoogle Scholar
  3. 3.
    Fayolle C, Deriaud E, Leclerc C (1991) In vivo induction of cytotoxic t cell response by a free synthetic peptide requires CD4+ T cell help. J Immunol 147:4069–4073PubMedGoogle Scholar
  4. 4.
    Bijker MS, van den Eeden SJ, Franken KL, Melief CJ, Offringa R, van der Burg SH (2007) CD8+ CTL priming by exact peptide epitopes in incomplete freund’s adjuvant induces a vanishing CTL response, whereas long peptides induce sustained CTL reactivity. J Immunol 179:5033–5040PubMedGoogle Scholar
  5. 5.
    Gritzapis AD, Voutsas IF, Lekka E, Papamichail M, Baxevanis CN (2010) Peptide vaccination breaks tolerance to HER-2/neu by generating vaccine-specific fasL+ CD4+ T cells: first evidence for intratumor apoptotic regulatory T cells. Cancer Res 70:2686–2696CrossRefPubMedGoogle Scholar
  6. 6.
    Kenter GG, Welters MJ, Valentijn AR, Lowik MJ, Berends-van der Meer DM, Vloon AP, Essahsah F, Fathers LM, Offringa R, Drijfhout JW, Wafelman AR, Oostendorp J, Fleuren GJ, van der Burg SH, Melief CJ (2009) Vaccination against HPV-16 oncoproteins for vulvar intraepithelial neoplasia. N Engl J Med 361:1838–1847CrossRefPubMedGoogle Scholar
  7. 7.
    Godet Y, Moreau-Aubry A, Guilloux Y, Vignard V, Khammari A, Dreno B, Jotereau F, Labarriere N (2008) Meloe-1 is a new antigen overexpressed in melanomas and involved in adoptive T cell transfer efficiency. J Exp Med 205:2673–2682CrossRefPubMedGoogle Scholar
  8. 8.
    Godet Y, Moreau-Aubry A, Mompelat D, Vignard V, Khammari A, Dreno B, Lang F, Jotereau F, Labarriere N (2010) An additional ORF on meloe cDNA encodes a new melanoma antigen, MELOE-2, recognized by melanoma-specific T cells in the HLA-A2 context. Cancer Immunol Immunother 59:431–439CrossRefPubMedGoogle Scholar
  9. 9.
    Godet Y, Desfrancois J, Vignard V, Schadendorf D, Khammari A, Dreno B, Jotereau F, Labarriere N (2010) Frequent occurrence of high affinity t cells against meloe-1 makes this antigen an attractive target for melanoma immunotherapy. Eur J Immunol 40:1786–1794CrossRefPubMedGoogle Scholar
  10. 10.
    Gervois N, Labarriere N, Le Guiner S, Pandolfino MC, Fonteneau JF, Guilloux Y, Diez E, Dreno B, Jotereau F (2000) High avidity melanoma-reactive cytotoxic T lymphocytes are efficiently induced from peripheral blood lymphocytes on stimulation by peptide-pulsed melanoma cells. Clin Cancer Res 6:1459–1467PubMedGoogle Scholar
  11. 11.
    Davodeau F, Difilippantonio M, Roldan E, Malissen M, Casanova JL, Couedel C, Morcet JF, Merkenschlager M, Nussenzweig A, Bonneville M, Malissen B (2001) The tight interallelic positional coincidence that distinguishes t-cell receptor jalpha usage does not result from homologous chromosomal pairing during valphajalpha rearrangement. EMBO J 20:4717–4729CrossRefPubMedGoogle Scholar
  12. 12.
    Arden B, Clark SP, Kabelitz D, Mak TW (1995) Human t-cell receptor variable gene segment families. Immunogenetics 42:455–500PubMedGoogle Scholar
  13. 13.
    Espevik T, Nissen-Meyer J (1986) A highly sensitive cell line, WEHI 164 clone 13, for measuring cytotoxic factor/tumor necrosis factor from human monocytes. J Immunol Methods 95:99–105CrossRefPubMedGoogle Scholar
  14. 14.
    Faure F, Mantegazza A, Sadaka C, Sedlik C, Jotereau F, Amigorena S (2009) Long-lasting cross-presentation of tumor antigen in human dc. Eur J Immunol 39:380–390CrossRefPubMedGoogle Scholar
  15. 15.
    Acosta-Rodriguez EV, Rivino L, Geginat J, Jarrossay D, Gattorno M, Lanzavecchia A, Sallusto F, Napolitani G (2007) Surface phenotype and antigenic specificity of human interleukin 17-producing t helper memory cells. Nat Immunol 8:639–646CrossRefPubMedGoogle Scholar
  16. 16.
    Hunder NN, Wallen H, Cao J, Hendricks DW, Reilly JZ, Rodmyre R, Jungbluth A, Gnjatic S, Thompson JA, Yee C (2008) Treatment of metastatic melanoma with autologous CD4+ T cells against NY-ESO-1. N Engl J Med 358:2698–2703CrossRefPubMedGoogle Scholar
  17. 17.
    Perez-Diez A, Joncker NT, Choi K, Chan WF, Anderson CC, Lantz O, Matzinger P (2007) CD4 cells can be more efficient at tumor rejection than CD8 cells. Blood 109:5346–5354CrossRefPubMedGoogle Scholar
  18. 18.
    Godefroy E, Scotto L, Souleimanian NE, Ritter G, Old LJ, Jotereau F, Valmori D, Ayyoub M (2006) Identification of two melan-a CD4+ T cell epitopes presented by frequently expressed MHC class II alleles. Clin Immunol 121:54–62CrossRefPubMedGoogle Scholar
  19. 19.
    Jager E, Jager D, Karbach J, Chen YT, Ritter G, Nagata Y, Gnjatic S, Stockert E, Arand M, Old LJ, Knuth A (2000) Identification of NY-ESO-1 epitopes presented by human histocompatibility antigen (HLA)-DRB4*0101–0103 and recognized by CD4(+) T lymphocytes of patients with NY-ESO-1-expressing melanoma. J Exp Med 191:625–630CrossRefPubMedGoogle Scholar
  20. 20.
    Schultz ES, Lethe B, Cambiaso CL, Van Snick J, Chaux P, Corthals J, Heirman C, Thielemans K, Boon T, van der Bruggen P (2000) A MAGE-A3 peptide presented by HLA-DP4 is recognized on tumor cells by CD4+ cytolytic t lymphocytes. Cancer Res 60:6272–6275PubMedGoogle Scholar
  21. 21.
    Wang XF, Kerzerho J, Adotevi O, Nuyttens H, Badoual C, Munier G, Oudard S, Tu S, Tartour E, Maillere B (2008) Comprehensive analysis of HLA-DR- and HLA-DP4-restricted CD4+ T cell response specific for the tumor-shared antigen survivin in healthy donors and cancer patients. J Immunol 181:431–439PubMedGoogle Scholar
  22. 22.
    Kobayashi H, Song Y, Hoon DS, Appella E, Celis E (2001) Tumor-reactive t helper lymphocytes recognize a promiscuous MAGE-A3 epitope presented by various major histocompatibility complex class II alleles. Cancer Res 61:4773–4778PubMedGoogle Scholar
  23. 23.
    Kobayashi H, Lu J, Celis E (2001) Identification of helper t-cell epitopes that encompass or lie proximal to cytotoxic t-cell epitopes in the gp100 melanoma tumor antigen. Cancer Res 61:7577–7584PubMedGoogle Scholar
  24. 24.
    Chicz RM, Urban RG, Gorga JC, Vignali DA, Lane WS, Strominger JL (1993) Specificity and promiscuity among naturally processed peptides bound to HLA-DR alleles. J Exp Med 178:27–47CrossRefPubMedGoogle Scholar
  25. 25.
    Kennedy R, Celis E (2008) Multiple roles for CD4+ t cells in anti-tumor immune responses. Immunol Rev 222:129–144CrossRefPubMedGoogle Scholar
  26. 26.
    Bioley G, Jandus C, Tuyaerts S, Rimoldi D, Kwok WW, Speiser DE, Tiercy JM, Thielemans K, Cerottini JC, Romero P (2006) Melan-a/mart-1-specific cd4 T cells in melanoma patients: identification of new epitopes and ex vivo visualization of specific t cells by MHC class II tetramers. J Immunol 177:6769–6779PubMedGoogle Scholar
  27. 27.
    Manici S, Sturniolo T, Imro MA, Hammer J, Sinigaglia F, Noppen C, Spagnoli G, Mazzi B, Bellone M, Dellabona P, Protti MP (1999) Melanoma cells present a MAGE-3 epitope to CD4(+) cytotoxic t cells in association with histocompatibility leukocyte antigen DR11. J Exp Med 189:871–876CrossRefPubMedGoogle Scholar
  28. 28.
    Chaux P, Vantomme V, Stroobant V, Thielemans K, Corthals J, Luiten R, Eggermont AM, Boon T, van der Bruggen P (1999) Identification of mage-3 epitopes presented by HLA-DR molecules to CD4(+) T lymphocytes. J Exp Med 189:767–778CrossRefPubMedGoogle Scholar
  29. 29.
    Rammensee H, Bachmann J, Emmerich NP, Bachor OA, Stevanovic S (1999) Syfpeithi: database for MHC ligands and peptide motifs. Immunogenetics 50:213–219CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Anne Rogel
    • 1
  • Virginie Vignard
    • 1
    • 2
  • Mathilde Bobinet
    • 1
  • Nathalie Labarriere
    • 1
  • François Lang
    • 1
    • 3
  1. 1.INSERM U892–CRCNA, IRTUNNantes cedex 1France
  2. 2.CHU of NantesNantesFrance
  3. 3.Pharmacology Department, UFR des Sciences PharmaceutiquesUniversité de NantesNantesFrance

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