Cancer Immunology, Immunotherapy

, Volume 53, Issue 2, pp 125–134 | Cite as

Phase I/II study of treatment with dendritic cell vaccines in patients with disseminated melanoma

  • Peter HerseyEmail author
  • Scott W. Menzies
  • Gary M. Halliday
  • Tam Nguyen
  • Margaret L. Farrelly
  • Chitra DeSilva
  • Margaret Lett
Original Article


Previous studies have suggested that immunotherapy with dendritic cell (DC) vaccines may be effective in treatment of patients with AJCC stage IV melanoma. We examined this treatment in phase I/II studies in 33 patients with good performance status and low volume disease. Nineteen patients received DCs plus autologous lysates and 14 patients DCs plus peptides from the melanoma antigens MAGE-3.A2, tyrosinase, gp100, and MART-1. Keyhole limpet hemocyanin (KLH) was used as a helper protein and influenza peptide was given as a positive control. DCs were produced from adherent cells in blood lymphocytes (monocytic DCs), grown in IL-4 and GM-CSF without a maturation step. The DCs were injected into inguinal lymph nodes at weekly intervals (×4), 2 weeks (×1), and 4-weekly intervals (×2). There were 3 responses (3 partial responses) and 1 mixed response in the 19 patients treated with DCs plus autologous lysates. No responses were seen in the group treated with DCs plus peptides. Stable disease (defined as no progression over a period of 3 months) was seen in 4 patients in group 1 and 5 patients in group 2. Treatment was not associated with significant side effects. We examined whether DTH skin tests or assays of IFN-γ cytokine production may be useful predictors of clinical responses. Twenty-two of 30 patients had DTH responses to KLH and 12 of 13 patients had DTH responses to the influenza peptide. Five of 15 DTH responses were seen against autologous lysates. This was strongly correlated with clinical responses. Approximately half the patients had responses to MART-1 peptide and a third to the other melanoma peptides. Similarly, cytokine production assays showed responses to influenza in 7 of 13 patients, and approximately one third of patients had responses to the other peptides. No IFN-γ responses were seen in 5 patients against their autologous lysates. There was no correlation between assays of IFN-γ production and clinical responses. The present studies suggest that autologous lysates may be more effective than the melanoma peptides used in the study as the source of antigen for DC vaccines. DTH responses to autologous lysates appear useful predictors of clinical responses, but further work is needed to identify other measures associated with clinical responses.


Melanoma Dendritic cells Melanoma peptides Clinical responses Immune responses 



dendritic cells


delayed hypersensitivity skin tests


keyhole limpet hemocyanin


cytotoxic T lymphocytes



This work was supported by the National Health and Medical Research Council of Australia and in part by the Sydney Melanoma Foundation and the Hunter Melanoma Foundation.


  1. 1.
    Atkins MB (1997) The treatment of metastatic melanoma with chemotherapy: biologics. Curr Opin Oncol 9:205PubMedGoogle Scholar
  2. 2.
    Von Eschen KB, Mitchell MS (1997) Phase III trial of melacine melanoma theraccine versus combination chemotherapy in the treatment of stage IV melanoma. Melanoma Res 7:S51PubMedGoogle Scholar
  3. 3.
    Hersey P (2002) Advances in the non-surgical treatment of melanoma. Expert Opin Investig Drugs 11:75PubMedGoogle Scholar
  4. 4.
    Steinman RM (1996) Dendritic cells and immune-based therapies. Exp Hematol 24:859PubMedGoogle Scholar
  5. 5.
    Banchereau J, Paczesny S, Blanco P, Bennett L, Pascual V, Fay J, Palucka AK (2003) Dendritic cells: controllers of the immune system and a new promise for immunotherapy. Ann N Y Acad Sci 987:180PubMedGoogle Scholar
  6. 6.
    Grabbe S, Bruvers S, Gallo RL, Knisely TL, Nazareno R, Granstein RD (1991) Tumor antigen presentation by murine epidermal cells. J Immunol 146:3656PubMedGoogle Scholar
  7. 7.
    Cavanagh LL, Sluyter R, Henderson KG, Barnetson RS, Halliday GM (1996) Epidermal Langerhans’ cell induction of immunity against an ultraviolet-induced skin tumour. Immunology 87:475PubMedGoogle Scholar
  8. 8.
    Mayordomo JI, Zorina T, Storkus WJ, Zitvogel L, Celluzzi C, Falo LD, Melief CJ, Ildstad ST, Kast WM, Deleo AB (1995) Bone marrow-derived dendritic cells pulsed with synthetic tumour peptides elicit protective and therapeutic antitumour immunity. Nat Med 1:1297PubMedGoogle Scholar
  9. 9.
    Celluzzi CM, Mayordomo JI, Storkus WJ, Lotze MT, Falo LD Jr (1996) Peptide-pulsed dendritic cells induce antigen-specific CTL-mediated protective tumor immunity. J Exp Med 183:283PubMedGoogle Scholar
  10. 10.
    Zitvogel L, Mayordomo JI, Tjandrawan T, Deleo AB, Clarke MR, Lotze MT, Storkus WJ (1996) Therapy of murine tumors with tumor peptide-pulsed dendritic cells: dependence on T cells, B7 costimulation, and T helper cell 1-associated cytokines. J Exp Med 183:87PubMedGoogle Scholar
  11. 11.
    Nestle FO, Alijagic S, Gilliet M, Sun Y, Grabbe S, Dummer R, Burg G, Schadendorf D (1998) Vaccination of melanoma patients with peptide- or tumor lysate-pulsed dendritic cells. Nat Med 4:328PubMedGoogle Scholar
  12. 12.
    Thurner B, Haendle I, Roder C, Dieckmann D, Keikavoussi P, Jonuleit H, Bender A, Maczek C, Schreiner D, van Den Driesch P, Brocker EB, Steinman RM, Enk A, Kampgen E, Schuler G (1999) Vaccination with Mage-3A1 peptide-pulsed mature, monocyte-derived dendritic cells expands specific cytotoxic T cells and induces regression of some metastases in advanced stage IV melanoma. J Exp Med 190:1669PubMedGoogle Scholar
  13. 13.
    Mackensen A, Herbst B, Chen JL, Kohler G, Noppen C, Herr W, Spagnoli GC, Cerundolo V, Lindemann A (2000) 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 86:385PubMedGoogle Scholar
  14. 14.
    Panelli MC, Wunderlich J, Jeffries J, Wang E, Mixon A, Rosenberg SA, Marincola FM (2000) Phase I study in patients with metastatic melanoma of immunization with dendritic cells presenting epitopes derived from the melanoma-associated antigens MART-1 and gp100. J Immunother 23:487PubMedGoogle Scholar
  15. 15.
    Chakraborty NG, Sporn JR, Tortora AF, Kurtzman SH, Yamase HY, Ergin MT, Mukherji B (1998) Immunization with a tumor-cell-lysate-loaded autologous-antigen-presenting-cell-based vaccine in melanoma. Cancer Immunol Immunother 47:58CrossRefPubMedGoogle Scholar
  16. 16.
    Banchereau J, Palucka AK, Dhodapkar M, Burkeholder S, Taquet N, Rolland A, Taquet S, Coquery S, Wittkowski KM, Bhardwaj N, Pineiro L, Steinman R, Fay J (2001) Immune and clinical responses in patients with metastatic melanoma to CD34(+) progenitor-derived dendritic cell vaccine. Cancer Res 61:6451PubMedGoogle Scholar
  17. 17.
    Therasse P, Arbuck SG, Eisenhauer EA, Wanders J, Kaplan RS, Rubinstein L, Verweij J, Van Glabbeke M, van Oosterom AT, Christian MC, Gwyther SG (2000) New guidelines to evaluate the response to treatment in solid tumors. J Nat Cancer Inst 92:205CrossRefPubMedGoogle Scholar
  18. 18.
    Hersey P (1997) Melanoma vaccines: prospects for the treatment of melanoma. Expert Opin Investig Drugs 6:267Google Scholar
  19. 19.
    Valmori D, Fonteneau J-F, Lizana CM, Gervois N, Lienard D, Rimoldi D, Jongeneel V, Jotereau F, Cerottini J-C, Romero P (1998) Enhanced generation of specific tumor-reactive CTL in vitro by selected Melan-A/MART-1 immunodominant peptide analogues. J Immunol 160:1750PubMedGoogle Scholar
  20. 20.
    Parkhurst MR, Salgaller ML, Southwood S, Robbins PF, Sette A, Rosenberg SA, Kawakami Y (1996) Improved induction of melanoma-reactive CTL with peptides from the melanoma antigen gp100 modified at HLA-A*0201-binding residues. J Immunol 157:2539PubMedGoogle Scholar
  21. 21.
    Skipper JCA, Hendrickson RC, Gulden PH, Brichard V, van Pel A, Chen Y, Shabanowitz J, Wolfel T, Slingluff CLJr, Boon T, Hunt DF, Engelhard VH (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 183:527PubMedGoogle Scholar
  22. 22.
    Jaeger E, Ringhoffer M, Dienes HP, Arand M, Karbach J, Jager D (1996) Granulocyte-macrophage-colony-stimulating factor enhances immune responses to melanoma-associated peptides in vivo. Int J Cancer 67:54PubMedGoogle Scholar
  23. 23.
    Saleh FH, Crotty KA, Hersey P, Menzies SW (2001) Primary melanoma tumour regression associated with an immune response to the tumour-assocated antigen melan-A/MART-1. Int J Cancer 94:551CrossRefPubMedGoogle Scholar
  24. 24.
    Nijman HW, Houbiers JGA, Vierboom MPM, Van Der Burg SH, Drijfhout JW, D’Amaro J, Kenemans P, Melief CJM, Kast WM (1993) Identification of peptide sequences that potentially trigger HLA-A2.1-restricted cytotoxic T lymphocytes. Eur J Immunol 23:1215PubMedGoogle Scholar
  25. 25.
    Salgaller ML, Marincola FM, Cormier JN, Rosenberg SA (1996) Immunization against epitopes in the human melanoma antigen gp100 following patient immunization with synthetic peptides. Cancer Res 56:4749PubMedGoogle Scholar
  26. 26.
    Cao X, Sugita M, Van Der Wel N, Lai J, Rogers RA, Peters PJ, Brenner MB (2002) CD1 molecules efficiently present antigen in immature dendritic cells and traffic independently of MHC class II during dendritic cell maturation. J Immunol 169:4770PubMedGoogle Scholar
  27. 27.
    Corbi AL, Lopez-Rodriguez C (1997) CD11c integrin gene promoter activity during myeloid differentation. Leuk Lymphoma 25:415PubMedGoogle Scholar
  28. 28.
    Smithers M, O’Connell K, MacFadyen S, Chambers M, Greenwood K, Boyce A, Abdul-Jabbar I, Barker K, Grimmett K, Walpole E, Thomas R (2003) Clinical response after intradermal immature dendritic cell vaccination in metastatic melanoma is associated with immune response to particulate antigen. Cancer Immunol Immunother 52:41PubMedGoogle Scholar
  29. 29.
    Lee K-H, Panelli MC, Kim CJ, Riker AI, Bettinotti MP, Roden MM, Fetsch P, Abati A, Rosenberg SA, Marincola FM (1998) Functional dissociation between local and systemic immune response during anti-melanoma peptide vaccination. J Immunol 161:4183PubMedGoogle Scholar
  30. 30.
    Clay TM, Hobeika AC, Mosca PJ, Lyerly HK, Morse MA (2001) Assays for monitoring cellular immune responses to active immunotherapy of cancer. Clin Cancer Res 7:1127PubMedGoogle Scholar
  31. 31.
    Whiteside TL, Zhao Y, Tsukishiro T, Elder EM, Gooding W, Baar J (2003) Enzyme-linked immunospot, cytokine flow cytometry, and tetramers in the detection of T-cell responses to a dendritic cell-based multipeptide vaccine in patients with melanoma. Clin Cancer Res 9:641PubMedGoogle Scholar
  32. 32.
    Yang S, Linette GP, Longerich S, Haluska FG (2002) Antimelanoma activity of CTL generated from peripheral blood mononuclear cells after stimulation with autologous dendritic cells pulsed with melanoma gp 100 peptide G209–2M is correlated to TCR avidity. J Immunol 169:531PubMedGoogle Scholar
  33. 33.
    Monsurro V, Nagorsen D, Wang E, Provenzano M, Dudley ME, Rosenberg SA, Marincola FM (2002) Functional heterogeneity of vaccine-induced CD8(+) T cells. J Immunol 168:5933PubMedGoogle Scholar
  34. 34.
    Morse MA, Clay TM, Mosca P, Lyerly HK (2002) Immunoregulatory T cells in cancer immunotherapy. Expert Opin Biol Ther 2:827PubMedGoogle Scholar
  35. 35.
    Javia LR, Rosenberg SA (2003) CD4+CD25+ suppressor lymphocytes in the circulation of patients immunized against melanoma antigens. J Immunother 26:85CrossRefPubMedGoogle Scholar
  36. 36.
    Zehntner S, Townsend W, Parkes J, Schmidt C, Down M, Bell J, Mulligan R, O’Rourke M, Ellem K, Thomas R (1999) Tumor metastasis biopsy as a surrogate marker of response to melanoma immunotherapy. Pathology 31:116CrossRefPubMedGoogle Scholar
  37. 37.
    Labeur MS, Roters B, Pers B, Mehling A, Luger TA, Schwarz T, Grabbe S (1999) Generation of tumor immunity by bone marrow-derived dendritic cells correlates with dendritic cell maturation. J Immunol 162:168PubMedGoogle Scholar
  38. 38.
    Jonuleit H, Giesecke-Tuettenberg A, Tuting T, Thurner-Schuler B, Stuge TB, Paragnik L, Kandemir A, Lee PP, Schuler G, Knop J, Enk AH (2001) A comparison of two types of dendritic cell as adjuvants for the induction of melanoma-specific T-cell responses in humans following intranodal injection. Int J Cancer 93:243PubMedGoogle Scholar
  39. 39.
    Jonuleit H, Giesecke A, Kandemir A, Paragnik L, Knop J, Enk AH (2000) Induction of tumor peptide-specific cytotoxic T cells under serum-free conditions by mature human dendritic cells. Arch Dermatol 292:325CrossRefPubMedGoogle Scholar
  40. 40.
    Adema GJ, de Vries IJM, Bernsen M, van Muijen GNP, Scharenborg NM, Lesterhuis WJ, Punt CJA, Figdor CG (2002) Immunological and clinical responses in melanoma patients after treatment with peptide-loaded dendritic cells administered via different routes of immunization. 7th symposium on dendritic cells, September 2002 (Abstract 019)Google Scholar
  41. 41.
    de Vries IJM, Krooshoop JEB, Scharenborg NM, Lesterhuis WJ, Diepstra JHS, van Muijen GNP, Strijk SP, Ruers TJ, Boerman OC, Oyen WJG, Adema GJ, Punt CJA, Figdor CG (2003) Effective migration of antigen-pulsed dendritic cells to lymph nodes in melanoma patients is determined by their maturation state. Cancer Res 63:12PubMedGoogle Scholar
  42. 42.
    Mahnke K, Schmitt E, Bonifaz L, Enk AH, Jonuleit H (2002) Immature, but not inactive: the tolerogenic function of immature dendritic cells. Immunol Cell Biol 80:477CrossRefPubMedGoogle Scholar
  43. 43.
    Dhodapkar MV, Steinman RM, Krasovsky J, Munz C, Bhardwaj N (2001) Antigen-specific inhibition of effector T cell function in humans after injection of immature dendritic cells. J Exp Med 193:233PubMedGoogle Scholar
  44. 44.
    Jonuleit H, Schmitt E, Schuler G, Knop J, Enk AH (2000) Induction of interleukin 10-producing, nonproliferating CD4(+) T cells with regulatory properties by repetitive stimulation with allogeneic immature human dendritic cells. J Exp Med 192:1213CrossRefPubMedGoogle Scholar
  45. 45.
    Roncarolo M-G, Levings MK, Traversari C (2001) Differentiation of T regulatory cells by immature dendritic cells. J Exp Med 193:F5CrossRefPubMedGoogle Scholar
  46. 46.
    Ranieri E, Kierstead LS, Zarour H, Kirkwood JM, Lotze MT, Whites T, Storkus WJ (2000) Dendritic cell/peptide cancer vaccines: clinical responsivene and epitope spreading. Immunol Invest 29:121PubMedGoogle Scholar
  47. 47.
    Shimizu K, Thomas EK, Giedlin M, Mule JJ (2001) Enhancement of tumor lysate- and peptide-pulsed dendritic cell-based vaccines by the addition of foreign helper protein. Cancer Res 61:2618PubMedGoogle Scholar
  48. 48.
    Millard AL, Ittelet D, Schooneman F, Bernard J (2003) Dendritic cell KLH loading requirements for efficient CD4(+) T-cell priming and help to peptide-specific cytotoxic T-cell response, in view of potential use in cancer vaccines. Vaccine 2:869CrossRefGoogle Scholar
  49. 49.
    Shimizu K, Fields RC, Giedlin M, Mule JJ (1999) Systemic administration of interleukin 2 enhances the therapeutic efficacy of dendritic cell-based tumor vaccines. Proc Natl Acad Sci U S A 96:2268CrossRefPubMedGoogle Scholar
  50. 50.
    Portielje JE, Gratama JW, van Ojik HH, Stoter G, Kruit WH (2003) IL-12: a promising adjuvant for cancer vaccination. Cancer Immunol Immunother 52:133PubMedGoogle Scholar
  51. 51.
    Asavaroengchai W, Kotera Y, Mule JJ (2002) Tumor lysate-pulsed dendritic cells can elicit an effective antitumor immune response during early lymphoid recovery. Proc Natl Acad Sci U S A 99:931CrossRefPubMedGoogle Scholar
  52. 52.
    Dudley ME, Wunderlich JR, Robbins PF, Yang JC, Hwu P, Schwartzentruber D, Topalian SL, Sherry R, Restifo NP, Hubicki AM, Robinson MR, Raffeld M, Dura P, Seipp CA, Rogers-Freezer L, Morton KE, Mavroukakis SA, White DE, Rosenberg SA (2002) Cancer regression and autoimmunity in patients after clonal repopulation with antitumor lymphocytes. Science 298:850CrossRefPubMedGoogle Scholar
  53. 53.
    Hersey P, Zhang XD (2001) How melanoma cells evade TRAIL-induced apoptosis. Nat Rev 1:142CrossRefGoogle Scholar
  54. 54.
    Hersey P, Zhang XD (2003) Overcoming resistance of cancer cells to apoptosis. J Cell Physiol 169:9CrossRefGoogle Scholar
  55. 55.
    Schuler-Thurner B, Schultz ES, Berger TG, Weinlich G, Ebner S, Woerl P, Bender A, Feuerstein B, Fritsch PO, Romani N, Schuler G (2002) Rapid induction of tumor-specific type 1 T helper cells in metastatic melanoma patients by vaccination with mature, cryopreserved, peptide-loaded monocyte-derived dendritic cells. J Exp Med 195:1279CrossRefPubMedGoogle Scholar
  56. 56.
    Gajewsky TF, Fallarino F, Vogelzang N, Posner MC, Ashikari A, Sherman ML (1999) Effective melanoma antigen vaccination without dendritic cells (DC): A phase I study of immunization with Mage3 or Melan-A peptide-pulsed autologous PBMC plus RhIL-12. Proc ASCO 18:539aGoogle Scholar
  57. 57.
    Chang AE, Redman BG, Whitfield JR, Nickoloff BJ, Braun TM, Lee PP, Geiger JD, Mule JJ (2002) A phase I trial of tumor lysate-pulsed dendritic cells in the treatment of advanced cancer. Clin Cancer Res 8:1021PubMedGoogle Scholar
  58. 58.
    Lau R, Wang F, Jeffery G, Marty V, Kuniyoshi J, Bade E, Ryback ME, Weber J (2001) Phase I trial of intravenous peptide-pulsed dendritic cells in patients with metastatic melanoma. J Immunother 24:66Google Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Peter Hersey
    • 1
    Email author
  • Scott W. Menzies
    • 2
  • Gary M. Halliday
    • 3
  • Tam Nguyen
    • 1
  • Margaret L. Farrelly
    • 1
  • Chitra DeSilva
    • 2
  • Margaret Lett
    • 2
  1. 1.Oncology and Immunology UnitNewcastleAustralia
  2. 2.Sydney Melanoma UnitRoyal Prince Alfred HospitalCamperdownAustralia
  3. 3.Department of DermatologyUniversity of SydneySydneyAustralia

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