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Immunologic hierarchy, class II MHC promiscuity, and epitope spreading of a melanoma helper peptide vaccine

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An Erratum to this article was published on 26 August 2014

Abstract

Immunization with a combination melanoma helper peptide (6MHP) vaccine has been shown to induce CD4+ T cell responses, which are associated with patient survival. In the present study, we define the relative immunogenicity and HLA allele promiscuity of individual helper peptides and identify helper peptide-mediated augmentation of specific CD8+ T cell responses. Thirty-seven participants with stage IIIB-IV melanoma were vaccinated with 6MHP in incomplete Freund’s adjuvant. The 6MHP vaccine is comprised of 6 peptides representing melanocytic differentiation proteins gp100, tyrosinase, Melan-A/MART-1, and cancer testis antigens from the MAGE family. CD4+ and CD8+ T cell responses were assessed in peripheral blood and in sentinel immunized nodes (SIN) by thymidine uptake after exposure to helper peptides and by direct interferon-γ ELIspot assay against 14 MHC class I-restricted peptides. Vaccine-induced CD4+ T cell responses to individual epitopes were detected in the SIN of 63 % (22/35) and in the peripheral blood of 38 % (14/37) of participants for an overall response rate of 65 % (24/37). The most frequently immunogenic peptides were MAGE-A3281–295 (49 %) and tyrosinase386–406 (32 %). Responses were not limited to HLA restrictions originally described. Vaccine-associated CD8+ T cell responses against class I-restricted peptides were observed in 45 % (5/11) of evaluable participants. The 6MHP vaccine induces both CD4+ and CD8+ T cell responses against melanoma antigens. CD4+ T cell responses were detected beyond reported HLA-DR restrictions. Induction of CD8+ T cell responses suggests epitope spreading and systemic activity mediated at the tumor site.

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Abbreviations

6MHP:

Combination melanoma helper peptide

CTL:

Cytotoxic T lymphocyte

PBMC:

Peripheral blood mononuclear cell

SI:

Stimulation index

SIN:

Sentinel immunized node

References

  1. Appay V, Zaunders JJ, Papagno L, Sutton J, Jaramillo A, Waters A et al (2002) Characterization of CD4(+) CTLs ex vivo. J Immunol 168:5954–5958

    Article  CAS  PubMed  Google Scholar 

  2. Greenberg PD, Kern DE, Cheever MA (1985) Therapy of disseminated murine leukemia with cyclophosphamide and immune lyt-1+, 2-T cells. Tumor eradication does not require participation of cytotoxic T cells. J Exp Med 161:1122–1134

    Article  CAS  PubMed  Google Scholar 

  3. Hunder NN, Wallen H, Cao J, Hendricks DW, Reilly JZ, Rodmyre R et al (2008) Treatment of metastatic melanoma with autologous CD4+ T cells against NY-ESO-1. N Engl J Med 358:2698–2703

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  4. Khong HT, Yang JC, Topalian SL, Sherry RM, Mavroukakis SA, White DE et al (2004) Immunization of HLA-A*0201 and/or HLA-DPbeta1*04 patients with metastatic melanoma using epitopes from the NY-ESO-1 antigen. J Immunother 27:472–477

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  5. Phan GQ, Touloukian CE, Yang JC, Restifo NP, Sherry RM, Hwu P et al (2003) Immunization of patients with metastatic melanoma using both class I- and class II-restricted peptides from melanoma-associated antigens. J Immunother 26:349–356

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  6. Wong R, Lau R, Chang J, Kuus-Reichel T, Brichard V, Bruck C et al (2004) Immune responses to a class II helper peptide epitope in patients with stage III/IV resected melanoma. Clin Cancer Res 10:5004–5013

    Article  CAS  PubMed  Google Scholar 

  7. Slingluff CL Jr, Petroni GR, Olson W, Czarkowski A, Grosh WW, Smolkin M et al (2008) Helper T-cell responses and clinical activity of a melanoma vaccine with multiple peptides from MAGE and melanocytic differentiation antigens. J Clin Oncol 26:4973–4980

    Article  PubMed Central  PubMed  Google Scholar 

  8. Slingluff CL Jr, Lee S, Zhao F, Chianese-Bullock KA, Olson WC, Butterfield LH et al (2013) A randomized phase II trial of multiepitope vaccination with melanoma peptides for cytotoxic T cells and helper T cells for patients with metastatic melanoma (E1602). Clin Cancer Res 19:4228–4238

    Article  CAS  PubMed  Google Scholar 

  9. Panina-Bordignon P, Tan A, Termijtelen A, Demotz S, Corradin G, Lanzavecchia A (1989) Universally immunogenic T cell epitopes: promiscuous binding to human MHC class II and promiscuous recognition by T cells. Eur J Immunol 19:2237–2242

    Article  CAS  PubMed  Google Scholar 

  10. Slingluff CL Jr, Yamshchikov G, Neese P, Galavotti H, Eastham S, Engelhard VH et al (2001) Phase I trial of a melanoma vaccine with gp100(280-288) peptide and tetanus helper peptide in adjuvant: immunologic and clinical outcomes. Clin Cancer Res 7:3012–3024

    CAS  PubMed  Google Scholar 

  11. Johnson RP, Trocha A, Yang L, Mazzara GP, Panicali DL, Buchanan TM et al (1991) HIV-1 gag-specific cytotoxic T lymphocytes recognize multiple highly conserved epitopes. Fine specificity of the gag-specific response defined by using unstimulated peripheral blood mononuclear cells and cloned effector cells. J Immunol 147:1512–1521

    CAS  PubMed  Google Scholar 

  12. Zarour HM, Kirkwood JM, Kierstead LS, Herr W, Brusic V, Slingluff CL Jr et al (2000) Melan-A/MART-1(51-73) represents an immunogenic HLA-DR4-restricted epitope recognized by melanoma-reactive CD4(+) T cells. Proc Natl Acad Sci USA 97:400–405

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  13. Topalian SL, Gonzales MI, Parkhurst M, Li YF, Southwood S, Sette A et al (1996) Melanoma-specific CD4+ T cells recognize nonmutated HLA-DR-restricted tyrosinase epitopes. J Exp Med 183:1965–1971

    Article  CAS  PubMed  Google Scholar 

  14. Kobayashi H, Kokubo T, Sato K, Kimura S, Asano K, Takahashi H et al (1998) CD4+ T cells from peripheral blood of a melanoma patient recognize peptides derived from nonmutated tyrosinase. Cancer Res 58:296–301

    CAS  PubMed  Google Scholar 

  15. Manici S, Sturniolo T, Imro MA, Hammer J, Sinigaglia F, Noppen C et al (1999) Melanoma cells present a MAGE-3 epitope to CD4(+) cytotoxic T cells in association with histocompatibility leukocyte antigen DR11. J Exp Med 189:871–876

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  16. Chaux P, Vantomme V, Stroobant V, Thielemans K, Corthals J, Luiten R et al (1999) Identification of MAGE-3 epitopes presented by HLA-DR molecules to CD4(+) T lymphocytes. J Exp Med 189:767–778

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  17. Halder T, Pawelec G, Kirkin AF, Zeuthen J, Meyer HE, Kun L et al (1997) Isolation of novel HLA-DR restricted potential tumor-associated antigens from the melanoma cell line FM3. Cancer Res 57:3238–3244

    CAS  PubMed  Google Scholar 

  18. Li K, Adibzadeh M, Halder T, Kalbacher H, Heinzel S, Muller C et al (1998) Tumour-specific MHC-class-II-restricted responses after in vitro sensitization to synthetic peptides corresponding to gp100 and annexin II eluted from melanoma cells. Cancer Immunol Immunother 47:32–38

    Article  CAS  PubMed  Google Scholar 

  19. Consogno G, Manici S, Facchinetti V, Bachi A, Hammer J, Conti-Fine BM et al (2003) Identification of immunodominant regions among promiscuous HLA-DR-restricted CD4+ T-cell epitopes on the tumor antigen MAGE-3. Blood 101:1038–1044

    Article  CAS  PubMed  Google Scholar 

  20. Benteyn D, Van Nuffel AM, Wilgenhof S, Corthals J, Heirman C, Neyns B et al (2013) Characterization of CD8+ T-cell responses in the peripheral blood and skin injection sites of melanoma patients treated with mRNA electroporated autologous dendritic cells (TriMixDC-MEL). Biomed Res Int 2013:976383

    PubMed Central  PubMed  Google Scholar 

  21. Wilgenhof S, Van Nuffel AM, Benteyn D, Corthals J, Aerts C, Heirman C et al (2013) A phase IB study on intravenous synthetic mRNA electroporated dendritic cell immunotherapy in pretreated advanced melanoma patients. Ann Oncol 24:2686–2693

    Article  CAS  PubMed  Google Scholar 

  22. Slingluff CL Jr, Petroni GR, Olson WC, Smolkin ME, Ross MI, Haas NB et al (2009) Effect of granulocyte/macrophage colony-stimulating factor on circulating CD8+ and CD4+ T-cell responses to a multipeptide melanoma vaccine: outcome of a multicenter randomized trial. Clin Cancer Res 15:7036–7044

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  23. Hailemichael Y, Dai Z, Jaffarzad N, Ye Y, Medina MA, Huang XF et al (2013) Persistent antigen at vaccination sites induces tumor-specific CD8(+) T cell sequestration, dysfunction and deletion. Nat Med 19:465–472

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  24. Salerno EP, Shea SM, Olson WC, Petroni GR, Smolkin ME, McSkimming C et al (2013) Activation, dysfunction and retention of T cells in vaccine sites after injection of incomplete freund’s adjuvant, with or without peptide. Cancer Immunol Immunother 62:1149–1159

    Article  CAS  PubMed  Google Scholar 

  25. 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 179:5033–5040

    Article  CAS  PubMed  Google Scholar 

  26. Chaux P, Luiten R, Demotte N, Vantomme V, Stroobant V, Traversari C et al (1999) Identification of five MAGE-A1 epitopes recognized by cytolytic T lymphocytes obtained by in vitro stimulation with dendritic cells transduced with MAGE-A1. J Immunol 163:2928–2936

    CAS  PubMed  Google Scholar 

  27. Kittlesen DJ, Thompson LW, Gulden PH, Skipper JC, Colella TA, Shabanowitz J et al (1998) Human melanoma patients recognize an HLA-A1-restricted CTL epitope from tyrosinase containing two cysteine residues: implications for tumor vaccine development. J Immunol 160:2099–2106

    CAS  PubMed  Google Scholar 

  28. Kawakami Y, Robbins PF, Wang X, Tupesis JP, Parkhurst MR, Kang X et al (1998) Identification of new melanoma epitopes on melanosomal proteins recognized by tumor infiltrating T lymphocytes restricted by HLA-A1, -A2, and -A3 alleles. J Immunol 161:6985–6992

    CAS  PubMed  Google Scholar 

  29. Traversari C, van der Bruggen P, Luescher IF, Lurquin C, Chomez P, Van Pel A et al (1992) A nonapeptide encoded by human gene MAGE-1 is recognized on HLA-A1 by cytolytic T lymphocytes directed against tumor antigen MZ2-E. J Exp Med 176:1453–1457

    Article  CAS  PubMed  Google Scholar 

  30. Gaugler B, Van den Eynde B, van der Bruggen P, Romero P, Gaforio JJ, De Plaen E et al (1994) Human gene MAGE-3 codes for an antigen recognized on a melanoma by autologous cytolytic T lymphocytes. J Exp Med 179:921–930

    Article  CAS  PubMed  Google Scholar 

  31. Celis E, Tsai V, Crimi C, DeMars R, Wentworth PA, Chesnut RW et al (1994) Induction of anti-tumor cytotoxic T lymphocytes in normal humans using primary cultures and synthetic peptide epitopes. Proc Natl Acad Sci USA 91:2105–2109

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  32. Skipper JC, Hendrickson RC, Gulden PH, Brichard V, Van Pel A, Chen Y et al (1996) An HLA-A2-restricted tyrosinase antigen on melanoma cells results from posttranslational modification and suggests a novel pathway for processing of membrane proteins. J Exp Med 183:527–534

    Article  CAS  PubMed  Google Scholar 

  33. Kawakami Y, Eliyahu S, Jennings C, Sakaguchi K, Kang X, Southwood S et al (1995) Recognition of multiple epitopes in the human melanoma antigen gp100 by tumor-infiltrating T lymphocytes associated with in vivo tumor regression. J Immunol 154:3961–3968

    CAS  PubMed  Google Scholar 

  34. Cox AL, Skipper J, Chen Y, Henderson RA, Darrow TL, Shabanowitz J et al (1994) Identification of a peptide recognized by five melanoma-specific human cytotoxic T cell lines. Science 264:716–719

    Article  CAS  PubMed  Google Scholar 

  35. Huang LQ, Brasseur F, Serrano A, De Plaen E, van der Bruggen P, Boon T et al (1999) Cytolytic T lymphocytes recognize an antigen encoded by MAGE-A10 on a human melanoma. J Immunol 162:6849–6854

    CAS  PubMed  Google Scholar 

  36. Kawakami Y, Eliyahu S, Sakaguchi K, Robbins PF, Rivoltini L, Yannelli JR et al (1994) Identification of the immunodominant peptides of the MART-1 human melanoma antigen recognized by the majority of HLA-A2-restricted tumor infiltrating lymphocytes. J Exp Med 180:347–352

    Article  CAS  PubMed  Google Scholar 

  37. Romero P, Dutoit V, Rubio-Godoy V, Lienard D, Speiser D, Guillaume P et al (2001) CD8+ T-cell response to NY-ESO-1: relative antigenicity and in vitro immunogenicity of natural and analogue sequences. Clin Cancer Res 7:766s–772s

    CAS  PubMed  Google Scholar 

  38. Skipper JC, Kittlesen DJ, Hendrickson RC, Deacon DD, Harthun NL, Wagner SN et al (1996) Shared epitopes for HLA-A3-restricted melanoma-reactive human CTL include a naturally processed epitope from pmel-17/gp100. J Immunol 157:5027–5033

    CAS  PubMed  Google Scholar 

  39. Hogan KT, Sutton JN, Chu KU, Busby JA, Shabanowitz J, Hunt DF et al (2005) Use of selected reaction monitoring mass spectrometry for the detection of specific MHC class I peptide antigens on A3 supertype family members. Cancer Immunol Immunother 54:359–371

    Article  CAS  PubMed  Google Scholar 

  40. Wang RF, Johnston SL, Zeng G, Topalian SL, Schwartzentruber DJ, Rosenberg SA (1998) A breast and melanoma-shared tumor antigen: T cell responses to antigenic peptides translated from different open reading frames. J Immunol 161:3598–3606

    CAS  PubMed  Google Scholar 

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Surgery/Division of Surgical Oncology, University of Virginia Health System, PO Box 800679, Charlottesville, VA, 22908-0679, USA

    Yinin Hu, Walter C. Olson, Kimberly A. Chianese-Bullock & Craig L. Slingluff Jr.

  2. Department of Public Health Sciences, University of Virginia Health System, Charlottesville, VA, 22908, USA

    Gina R. Petroni & Mark E. Smolkin

  3. Cancer Center, University of Virginia Health System, Charlottesville, VA, 22908, USA

    Andrea Czarkowski

  4. Department of Medicine/Division of Hematology-Oncology, University of Virginia Health System, Charlottesville, VA, 22908, USA

    William W. Grosh

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  1. Yinin Hu
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  2. Gina R. Petroni
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  3. Walter C. Olson
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  8. Craig L. Slingluff Jr.
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Correspondence to Yinin Hu.

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Hu, Y., Petroni, G.R., Olson, W.C. et al. Immunologic hierarchy, class II MHC promiscuity, and epitope spreading of a melanoma helper peptide vaccine. Cancer Immunol Immunother 63, 779–786 (2014). https://doi.org/10.1007/s00262-014-1551-x

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  • DOI: https://doi.org/10.1007/s00262-014-1551-x

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