A phase II trial of vaccination with autologous, tumor-derived heat-shock protein peptide complexes Gp96, in combination with GM-CSF and interferon-α in metastatic melanoma patients

  • Lorenzo Pilla
  • Roberto Patuzzo
  • Licia Rivoltini
  • Michele Maio
  • Elisabetta Pennacchioli
  • Elda Lamaj
  • Andrea Maurichi
  • Samuele Massarut
  • Alfonso Marchianò
  • Cristina Santantonio
  • Diego Tosi
  • Flavio Arienti
  • Agata Cova
  • Gloria Sovena
  • Adriano Piris
  • Daisuke Nonaka
  • Ilaria Bersani
  • Annabella Di Florio
  • Mariani Luigi
  • Pramod K. Srivastava
  • Axel Hoos
  • Mario Santinami
  • Giorgio Parmiani
Original Article

Abstract

The aim of this study was to determine the immunogenicity and antitumor activity of autologous, tumor-derived heat shock protein gp96-peptide complex vaccine (HSPPC-96; Oncophage®) given with GM-CSF and IFN-α in pre-treated metastatic (AJCC stage IV) melanoma patients. Patients underwent surgical resection of metastatic lesions for HSPPC-96 production. HSPPC-96 was administered subcutaneously (s.c.) in four weekly intervals (first cycle). Patients with more available vaccine and absence of progressive disease received four additional injections in 2-week intervals (second cycle) or more. GM-CSF was given s.c. at the same site at days –1, 0 and +1, while IFN-α (3 MU) was administered s.c. at a different site at days +4 and +6. Antigen-specific anti-melanoma T and NK lymphocyte response was assessed by enzyme-linked immunospot assay on peripheral blood mononuclear cells obtained before and after vaccination. Thirty-eight patients were enrolled, 20 received at least four injections (one cycle) of HSPPC-96 and were considered assessable. Toxicity was mild and most treatment-related adverse events were local erythema and induration at the injection site. Patients receiving at least four injections of HSPPC-96 were considered evaluable for clinical response: of the 18 patients with measurable disease post surgery, 11 showed stable disease (SD). The ELISPOT assay revealed an increased class I HLA-restricted T and NK cell-mediated post-vaccination response in 5 out of 17 and 12 out of the 18 patients tested, respectively. Four of the five class I HLA-restricted T cell responses fall in the group of SD patients. Vaccination with autologous HSPPC-96 together with GM-CSF and IFN-α is feasible and accompanied by mild local and systemic toxicity. Both tumor-specific T cell-mediated and NK cell responses were generated in a proportion of patients. Clinical activity was limited to SD. However, both immunological and clinical responses were not improved as compared with those recorded in a previous study investigating HSPPC-96 monotherapy.

Keywords

Vaccination Metastatic melanoma Heat shock proteins GM-CSF IFN-α Phase II trial 

Abbreviations

AEs

adverse events

DTH

delayed-type hypersensitivity

ELISPOT

enzyme-linked immunospot

GM-CSF

granulocyte/macrophage-colony stimulating factor

HSP

heat shock proteins

HSPPC-96

heat shock proteins peptide complexes gp96

IFN

interferon

IHC

immunohistochemistry

IL

interleukin

NK

natural killer

PBMC

peripheral blood mononuclear cells

RECIST

response evaluation criteria in solid tumors

s.c.

subcutaneously

References

  1. 1.
    Belli F, Testori A, Rivoltini L, Maio M, Andreola G, Sertoli MR, Gallino P, Piris A, Cattelan A, Lazzari I, Carrabba M, Scita G et al (2002) Vaccination of metastatic melanoma patients with autologous tumor-derived heat shock protein gp96-peptide complexes: Clinical and immunologic findings. J Clin Oncol 20:4169–4180PubMedCrossRefGoogle Scholar
  2. 2.
    Parmiani G, Testori A, Maio M, Castelli C, Rivoltini L, Pilla L, Belli F, Mazzaferro V, Coppa J, Patuzzo R, Sertoli MR, Hoos A et al (2004) Heat-shock proteins and their use as anti-cancer vaccines. Clin Cancer Res 10:8142–8146PubMedCrossRefGoogle Scholar
  3. 3.
    Inaba K, Inaba M, Romani N, Aya H, Deguchi M, Ikehara S, Muramatsu S, Steinman RM (1992) Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte-macrophage colony stimulating factor. J Exp Med 176:1693–1699PubMedCrossRefGoogle Scholar
  4. 4.
    Soiffer R, Lynch T, Mihm M, Jung K, Rhuda C, Schmollinger JC, Hodi FS, Liebster L, Lam P, Mentzer S, Singer S, Tanabe KK et al (1998) Vaccination with irradiated autologous melanoma cells engineered to secrete human granulocyte-macrophage-colony stimulating factor generates potent antitumor immunity in patients with metastatic melanoma. Proc Natl Acad Sci USA 95:13141–13146PubMedCrossRefGoogle Scholar
  5. 5.
    Weber J, Sondak VK, Scotland R, Phillip R, Wang F, Rubio V, Stuge TB, Groshen SG, Gee C, Jeffery GG, Sian S, Lee PP (2003) Granulocyte-macrophage-colony stimulating factor added to a multipeptide vaccine for resected stage II melanoma. Cancer 97:186–200PubMedCrossRefGoogle Scholar
  6. 6.
    Marincola FM, Jaffee EM, Hicklin DJ, Ferrone S (2000) Escape of human solid tumors from T cell recognition: molecular mechanisms and functional significance. Adv Immunol 74:181–273PubMedCrossRefGoogle Scholar
  7. 7.
    Giacomini M, Fraioli R, Calabro AM, DiFilippo F, Natali PG (1991) Class I major histocompatibility complex enhancement by recombinant leukocyte interferon in the peripheral blood mononuclear cells and plasma of melanoma patients. Cancer Res 51:652–656PubMedGoogle Scholar
  8. 8.
    Therasse P, Arbuck SG, Eisenhauer EA (2000) New guidelines to evaluate the response to treatment in solid tumors. European Organization for Research and Treatment of Cancer, National Cancer Institute of the United States, National Cancer Institute of Canada. J Natl Cancer Inst 92:205–216PubMedCrossRefGoogle Scholar
  9. 9.
    Mazzaferro V, Coppa J, Carrabba MG, Rivoltini L, Schiavo M, Regalia E, Mariani L, Camerini T, Marchianò A, Andreola S, Camerini R, Corsi M et al (2003) Vaccination with autologous tumor-derived heat-shock protein Gp96 after liver resection for metastatic colorectal cancer. Clin Cancer Res 9:3235–3245PubMedGoogle Scholar
  10. 10.
    Pass HA, Schwarz SL, Wunderlich JR, Rosenberg SA (1998) Immunization of patients with melanoma peptide vaccines: Immunologic assessment using the ELISPOT assay. Cancer J Sci Am 4:316–323PubMedGoogle Scholar
  11. 11.
    Lewis JJ, Janetzki S, Schaed S, Panageas KS, Wang S, Williams L, Meyers M, Butterworth L, Livingston PO, Chapman PB, Houghton AN (2000) Evaluation of CD8+ T-cell frequency by the ELISPOT assay in healthy individuals and in patients with metastatic melanoma immunized with tyrosinase peptide. Int J Cancer 87:391–398PubMedCrossRefGoogle Scholar
  12. 12.
    Scheibenbogen C, Romero P, Rivoltini L, Herr W, Schmittel A, Cerottini JC, Woelfel T, Eggermont AMM, Keilholz U (2000) Quantification of antigen-reactive T cells in peripheral blood by IFN-γ-ELISPOT and chromium release assay: a four centre comparative trial. J Immunol Methods 244:81–89PubMedCrossRefGoogle Scholar
  13. 13.
    Rivoltini L, Castelli C, Carrabba M, Mazzaferro V, Pilla L, Huber V, Coppa J, Gallino G, Scheibenbogen C, Squarcina P, Cova A, Camerini R et al (2003) Human tumor-derived heat shock protein 96 mediates in vitro activation and in vivo expansion of melanoma-and colon carcinoma-specific T cells. J Immunol 171:3467–3474PubMedGoogle Scholar
  14. 14.
    Kawakami Y, Eliyahu S, Delgado CH, Robbins PF, Rivoltini L, Topalian SL, Miki T, Rosenbergs SA (1994) Cloning of the gene coding for a shared human melanoma antigen recognized by autologous T cells infiltrating into tumor. Proc Natl Acad Sci USA 91:3515–3519PubMedCrossRefGoogle Scholar
  15. 15.
    Mihm MC Jr, Clemente CG, Cascinelli N (1996) Tumor infiltrating lymphocytes in lymph node melanoma metastases: a histopathologic prognostic indicator and an expression of local immune response. Lab Invest 74:43–47PubMedGoogle Scholar
  16. 16.
    Moretta L, Bottino C, Pende D, Mingari MC, Biassoni R, Moretta A (2002) Human natural killer cells: their origin, receptors and function. Eur J Immunol 32:1205–1211PubMedCrossRefGoogle Scholar
  17. 17.
    Tamura Y, Peng P, Liu K, Srivastava PK (1997) Immunotherapy of tumors with autologous tumor-derived heat shock protein preparations. Science 278:117–120PubMedCrossRefGoogle Scholar
  18. 18.
    Pilla L, Squarcina P, Coppa J, Mazzaferro V, Huber V, Pende D, Maccalli C, Sovena G, Mariani L, Castelli C, Parmiani G, Rivoltini L (2005) Natural killer and NK-Like T-cell activation in colorectal carcinoma patients treated with autologous tumor-derived heat shock protein 96. Cancer Res 65:3942–3949PubMedCrossRefGoogle Scholar
  19. 19.
    Janetzki S, Palla D, Rosenhauer V, Lochs H, Lewis JJ, Srivastava PK (2000) Immunization of cancer patients with autologous cancer-derived heat shock protein gp96 preparations: a pilot study. Int J Cancer 88:232–238PubMedCrossRefGoogle Scholar
  20. 20.
    Gastpar R, Gross C, Rossbacher L, Ellwart J, Riegger J, Multhoff G (2004) The cell surface-localized heat shock protein 70 epitope TKD induces migration and cytolytic activity selectively in human NK cells. J Immunol 172:972–980PubMedGoogle Scholar
  21. 21.
    Scheibenbogen C, Schmittel A, Keilhjolz U, Allgauer T, Hofmann U, Max R, Thiel E, Schadendorf D (2000) Phase 2 trial of vaccination with tyrosinase peptides and granulocyte-macrophage colony-stimulating factor in patients with metastatic melanoma. J Immunother 23:275–281PubMedCrossRefGoogle Scholar
  22. 22.
    Slingluff CL, Petroni GR, Yamshikov GV, Barnd DL, Eastham S, Galavotti H, Patterson JW, Deacon DH, Hibbitts S, Teates D, Neese PY, Grosh WW et al (2003) Clinical and immunological results of a randomized phase II trial of vaccination using four melanoma peptides either administered in granulocyte-macrophage colony stimulating factor in adjuvant or pulsed on dendritic cells. J Clin Oncol 21:4016–4026PubMedCrossRefGoogle Scholar
  23. 23.
    Ullenhag GJ, Frodin J-E, Mosolitis S, Kiaii S, Hassan M, Bonnet MC, Moingeon P, Mellstedt H, Rabbani H (2003) Immunization of colorectal carcinoma patients with a recombinant canarypox virus expressing the tumor antigen Ep-CAM/KSA (ALVAC-KSA) and granulocyte macrophage colony-stimulating factor induced a tumor-specific cellular immune response. Clin Cancer Res 9:2447–2456PubMedGoogle Scholar
  24. 24.
    Von Meheren M, Arlen P, Gulley J, Rogatko A, Cooper HS, Meropol NJ, Alpaugh RK, Davey M, McLaughlin S, Beard MT, Tsang KY, Schlom J, Weiner LM (2001) The influence of granulocyte macrophage colony-stimulating factor and prior chemotherapy on the immunological response to a vaccine (ALVAC-CEA B7.1) in patients with metastatic carcinoma. Clin Cancer Res 7:1181–1191Google Scholar
  25. 25.
    Dillman RO, Wiemann M, Nayak SK, deLeon C, Hood K, DePriest C (2003) Interferon-gamma or granulocyte-macrophage colony-stimulating factor administered as adjuvants with a vaccine of irradiated autologous tumor cells from short-term cell line cultures: a randomized phase 2 trial of the cancer biotherapy research group. J Immunother 26:367–373PubMedCrossRefGoogle Scholar
  26. 26.
    Pansky A, Hildebrand P, Fasler-Kan E, Beglinger C, Heim MH (2000) Defective JAK-STAT signal transduction pathway in melanoma cell resistant to growth inhibition by interferon-α. Int J Cancer 85:720–725PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Lorenzo Pilla
    • 1
  • Roberto Patuzzo
    • 2
  • Licia Rivoltini
    • 1
  • Michele Maio
    • 3
    • 10
  • Elisabetta Pennacchioli
    • 2
  • Elda Lamaj
    • 3
  • Andrea Maurichi
    • 2
  • Samuele Massarut
    • 3
  • Alfonso Marchianò
    • 4
  • Cristina Santantonio
    • 3
  • Diego Tosi
    • 1
  • Flavio Arienti
    • 5
  • Agata Cova
    • 1
  • Gloria Sovena
    • 1
  • Adriano Piris
    • 6
  • Daisuke Nonaka
    • 6
  • Ilaria Bersani
    • 1
  • Annabella Di Florio
    • 1
  • Mariani Luigi
    • 7
  • Pramod K. Srivastava
    • 8
  • Axel Hoos
    • 9
  • Mario Santinami
    • 2
  • Giorgio Parmiani
    • 1
  1. 1.Unit of Immunotherapy of Human TumorsIstituto Nazionale per lo Studio e la Cura dei TumoriMilanItaly
  2. 2.Unit of Melanoma and Sarcoma SurgeryIstituto Nazionale per lo Studio e la Cura dei TumoriMilanItaly
  3. 3.Centro di Riferimento OncologicoAvianoItaly
  4. 4.Unit of RadiologyIstituto Nazionale per lo Studio e la Cura dei TumoriMilanItaly
  5. 5.Unit of ImmunohematologyIstituto Nazionale per lo Studio e la Cura dei TumoriMilanItaly
  6. 6.Unit of PathologyIstituto Nazionale per lo Studio e la Cura dei TumoriMilanItaly
  7. 7.Unit of Medical Statistics and BiometryIstituto Nazionale per lo Studio e la Cura dei TumoriMilanItaly
  8. 8.Department of Microbiology and ImmunologyUniversity of ConnecticutFarmingtonUSA
  9. 9.Antigenics Inc.LexingtonUSA
  10. 10.Unit of Medical Oncologic and Immunotherapy, Department of OncologyAzienda Ospedaliera Universitaria SeneseSienaItaly

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