Journal of Neuro-Oncology

, Volume 78, Issue 1, pp 71–80 | Cite as

Technical hurdles in a pilot clinical trial of combined B7-2 and GM-CSF immunogene therapy for glioblastomas and melanomas

  • Ian F. Parney
  • Lung-Ji Chang
  • Maxine A. Farr-Jones
  • Chunhai Hao
  • Michael Smylie
  • Kenneth C. Petruk



Malignant glioblastomas and melanomas continue to have a dismal prognosis despite advances in conventional therapy. This has led to investigations of novel treatment strategies including immunogene therapy. We report a pilot clinical trial of combined B7-2 and GM-CSF immunogene therapy for gliomas and melanomas and discuss technical hurdles encountered.


Patients with recurrent malignant gliomas or medically refractory melanomas were vaccinated with irradiated autologous tumor cells transduced with B7-2 and GM-CSF genes using a retroviral vector. Patients were monitored for toxicity, inflammatory/immune reactions, and clinical status.


Vaccine preparation was attempted from 116 malignant glioma and 32 melanoma specimens. Adequate vaccines could only be prepared for five glioblastoma and three melanoma patients. Six patients (three recurrent glioblastomas and three melanomas) were actually vaccinated. Minor toxicities included flu-like symptoms (3/6), injection site erythema (4/6), and asymptomatic elevations in liver enzymes (3/6). Most patients showed evidence of an inflammatory response but specific anti-tumor immunity was not demonstrated. All six patients have died, although three patients with minimal residual disease at treatment had prolonged recurrence-free intervals after vaccination.


Combined B7-2 and GM-CSF immunogene therapy for glioblastomas and melanomas using autologous tumor cells has many technical pitfalls hindering large scale application and evaluation. As a result, this pilot study was too limited to draw meaningful conclusions regarding safety or anti-tumor immunity. While immunotherapy has been promising in pre-clinical studies, alternate strategies will be required to bring these benefits to patients.


costimulation cytokine gene therapy glioblastoma immunotherapy melanoma 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.



We would like to acknowledge the other members of the University of Alberta Cancer Immunogene Therapy Working Group (H. Vandenhoven RN, V. Urlacher BSc, K. Kane PhD, A. Gainer MD PhD, E. Solano MD, D. Fulton MD, and R. Urtasun MD) without whom this trial would have been impossible. This data was presented in part at the Congress of Neurological Surgeons, Philadelphia, September 2002. The University of Alberta Hospital Foundation, the Allard Family Foundation, the Eldon Foote Foundation, and the Alberta Heritage Foundation for Medical Research provided funds for this study. Dr. Parney is an Alberta Heritage Foundation for Medical Research Clinical Investigator.


  1. 1.
    Forsyth PAJ, Cairncross JG, Treatment of malignant glioma in adultsCurr Opin Neurol1995; 8:414–418PubMedCrossRefGoogle Scholar
  2. 2.
    Fulton D, Urtasun R, Forsyth, P, Phase II study of prolonged oral therapy with etoposide (VP16) for patients with recurrent malignant gliomaJ Neurooncol1996; 27(2):149–155CrossRefPubMedGoogle Scholar
  3. 3.
    Garrison M, Nathanson L, Prognosis and staging in melanomaSem Oncol; 23: 1996 725–733Google Scholar
  4. 4.
    Hancock BW, Harris S, Wheatley K, Gore M Adjuvant interferon-alpha in malignant melanoma: current statusCancer Treat Rev 2000; 26:81–89CrossRefPubMedGoogle Scholar
  5. 5.
    Antonia SJ, B7-1 gene-modified tumor cell vaccinesCurr Opin Mol Therap 1999; 1:50–56Google Scholar
  6. 6.
    Parney IF, Hao C, Petruk KC, Glioma immunology and immunotherapy: a reviewNeurosurgery2000; 46:778–792CrossRefPubMedGoogle Scholar
  7. 7.
    Dranoff G, Jaffee E, Lazenby A, et al. Vaccination with irradiated tumor cells engineered to secrete murine granulocyte-macrophage colony-stimulating factor stimulates potent, specific and long lasting anti-tumor immunityProc Natl Acad Sci USA 1993; 90:3539–3543PubMedCrossRefGoogle Scholar
  8. 8.
    Tahara H, Lotze MT, Antitumor effects of interleukin-12 (IL-12): applications for the immunotherapy and gene therapy of cancerGene Therapy1995; 2:96–106PubMedGoogle Scholar
  9. 9.
    Parney IF, Farr-Jones MA, Koshal A, Chang L-J, Petruk KC, Human brain tumor cell culture characterization following retrovirus-mediated immunostimulatory gene transferNeurosurgery 2002; 50:1094–1102CrossRefPubMedGoogle Scholar
  10. 10.
    Parney IF, Petruk KC, Zhang C, Farr-Jones MA, Sykes DB, Chang L-J, GM-CSF and B7-2 combination immunogene therapy in an allogeneic hu-PBL-SCID/beige mouse – human glioblastoma multiforme modelHum Gene Therapy1997; 8:1073–1085Google Scholar
  11. 11.
    Parney IF, Farr-Jones MA, Kane K, Chang L-J, Petruk KC, Human autologous in vitro models of glioma immunogene therapy using B7-2, GM-CSF, and IL12Can J Neurol Sci; 2002 29:267–275PubMedGoogle Scholar
  12. 12.
    Sampson JH, Archer GE, Ashley DM, et al. Subcutaneous vaccination with irradiated, cytokine-producing tumor cells stimulates CD8+ cell-mediated immunity against tumors located in the “immunologically privileged” central nervous systemProc Acad Sci USA1996; 93:10399–10404CrossRefGoogle Scholar
  13. 13.
    Yu JS, Burwick JA, Dranoff G, Breakefield XO, Gene therapy for metastatic brain tumors by vaccination with granulocyte-macrophage colony-stimulating factor-transduced tumor cellsHum Gene Therapy1997; 8:1065–1072Google Scholar
  14. 14.
    Ellem KAO, O’Rourke MGE, Johnson GR, et al. A case report: immune responses and clinical course of the first human use of granulocyte-macrophage colony-stimulating factor-transduced autologous melanoma cells for immunotherapyCancer Immunol Immunother; 1997 44:10–20CrossRefPubMedGoogle Scholar
  15. 15.
    De Gast GC, Gallee MPW, Spits H, et al. Immunological, pathological, and long-term clinical data of vaccination with autologous granulocyte-macrophage colony-stimulating factor-transduced tumor cells in metastatic melanomaCancer Gene Ther 2000; 7:1204Google Scholar
  16. 16.
    Robert Soiffer TL, Mihn M, Jung K, et al. Vaccination with irradiated autologous melanoma cells engineered to secrete human granulocyte-macrophage colony-stimulating factor generates potent antitumor immunity in patients with metastatic melanomaPNAS 1998; 95:13141–13146CrossRefPubMedGoogle Scholar
  17. 17.
    Simons JW, Jaffee EM, Weber CE, et al. Bioactivity of autologous irradiated renal cell carcinoma vaccines generated by ex vivo granulocyte-macrophage colony-stimulating factor gene transferCancer Res 1997; 57:1537–1546PubMedGoogle Scholar
  18. 18.
    Simons JW, Mikhak B, Chang J-F, et al. Induction of immunity to prostate cancer antigens: results of a clinical trial of vaccination with irradiated autologous prostate cells engineered to secrete granulocyte-macrophage colony-stimulating factor using ex vivo gene transferCancer Res1999; 59:195–204Google Scholar
  19. 19.
    Parney IF, Farr-Jones MA, Petruk KC, Improved technique for establishing short term human brain tumor culturesJ Neuro-Oncol; 1999 43:1–10CrossRefGoogle Scholar
  20. 20.
    Kohn E, Engraftment of gene-modified umbilical cord blood cells in neonates with adenosine deaminase deficiencyNature Med; 1995 1:1017–1023CrossRefPubMedGoogle Scholar
  21. 21.
    Robinson D, Elliott JF, Chang LJ, Retroviral vector with a CMV-IE/HIV-TAR hybrid LTR gives high basal expression levels and is upregulated by HIV-1 TatGene Therapy1995; 2:269–278PubMedGoogle Scholar
  22. 22.
    Miller AD, Buttimore C, Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus productionMol Cell Biol;1986 6:2895–2902PubMedGoogle Scholar
  23. 23.
    Miller AD, Miller DG, Garcia JV, Lynch CM, Use of retroviral vectors for gene transfer and expressionMeth Enzymol; 1993 217:581–599PubMedGoogle Scholar
  24. 24.
    Morford LA, Elliott LH, Carlson SL, Brooks WH, Roszman TL, T cell receptor-mediated signalling is defective in T cells obtained from patients with primary intracranial tumorsJ Immunol; 1997 159:4415–4425PubMedGoogle Scholar
  25. 25.
    Badie B, Schartner JM, Paul J, Bartley BA, Vorpahl J, Preston JK, Dexamethasone-induced abolition of the inflammatory response in an experimental glioma model: a flow cytometry studyJ Neurosurg 2000; 93:634–639PubMedCrossRefGoogle Scholar
  26. 26.
    Human gene marker/therapy clinical protocols (complete updated listing). Hum Gene Ther12: 2251–2337, 2001Google Scholar
  27. 27.
    Chang L-J, Urlacher V, Iwakuma T, Cui Y, Zucali J, Efficacy and safety analyses of a recombinant human immunodeficiency virus type 1 derived vector systemGene Therapy1999; 6:715–728CrossRefPubMedGoogle Scholar
  28. 28.
    Gainer AL, Young ATL, Parney IF, Petruk KC, Elliott JF, Gene-gun transfection of human glioma and melanoma cell lines with genes encoding human IL-12 and GM-CSFJ Neuro-Oncol; 2000 47:23–30CrossRefGoogle Scholar
  29. 29.
    Chang LJ, He J, Retroviral vectors for gene therapy of AIDS and cancerCurr Opin Mol Ther 2001; 3:468–475PubMedGoogle Scholar
  30. 30.
    Ashley DM, Sampson JH, Archer GE, Batra SK, Bigner DD, Hale LP, A genetically modified allogeneic cellular vaccine generates MHC class I-restricted cytotoxic responses against tumor-associated antigens and protects against CNS tumors in vivoJ Neuroimmunol; 1997 78:34–46CrossRefPubMedGoogle Scholar
  31. 31.
    Thomas MC, Greten TF, Pardoll DM, Jaffee EM, Enhanced tumor protection by granulocyte-macrophage colony-stimulating factor expression at the site of an allogeneic vaccineHum Gene Ther1998; 9:835–843PubMedCrossRefGoogle Scholar
  32. 32.
    Jaffee EM, Hruban RH, Biedrzycki B, et al. Novel allogeneic granulocyte-macrophage colony-stimulating factor-secreting tumor vaccine for pancreatic cancer: a phase I trial of safety and immune activationJ Clin Oncol 2001; 19:145–156PubMedGoogle Scholar
  33. 33.
    Roszman T, Elliott L, Brooks W, Modulation of T-cell function by gliomas Immunol Today1991; 12:370–374CrossRefPubMedGoogle Scholar
  34. 34.
    Sobol RE, Shawler DL, Carson C, et al. Interleukin 2 gene therapy of colorectal carcinoma with autologous irradiated tumor cells and genetically engineered fibroblasts: a phase 1 studyClin Cancer Res; 1999 5:2359–2365PubMedGoogle Scholar
  35. 35.
    Trudel S, Li Z, Dodgson C, et al. Adenovector engineered interleukin-2 expressing autologous plasma cell vaccination after high-dose chemotherapy for multiple myeloma – a phase 1 studyLeukemia2001; 15:846–854CrossRefPubMedGoogle Scholar
  36. 36.
    Dranoff G Interpreting cancer vaccine clinical trialsJ Gene Med1999; 1:80–83CrossRefPubMedGoogle Scholar
  37. 37.
    Altman J, Moss P, Goulder P, et al. Direct visualization and phenotypic analysis of virus-specific T-lymphocytes in HIV-infected individualsScience1996; 274:94–96CrossRefPubMedGoogle Scholar
  38. 38.
    Becker C, Pohla H, Frankenberger B, et al. Adoptive tumor therapy with T lymphocytes enriched through an interferon-gamma capture assayNature Med 2001; 7:1159–1162CrossRefPubMedGoogle Scholar
  39. 39.
    Parmiani G, Rodolfo M, Melani C, Immunological gene therapy with ex vivo gene-modified tumor cells: a critique and a reappraisalHum Gene Ther; 2000 11:1269–1275CrossRefPubMedGoogle Scholar
  40. 40.
    Chi DDJ, Merchant RE, Rand R, et al. Molecular detection of tumor-associated antigens shared by human cutaneous melanomas and gliomasAm J Pathol; 1997 150:2143–2152PubMedGoogle Scholar
  41. 41.
    Greenfield EA, Nguyen KA, Kuchroo VK, CD28/B7 costimulation: a reviewCrit Rev Immunol; 1998 18:389–418PubMedGoogle Scholar
  42. 42.
    Galea-Lauri J, Farzaneh F, Gaken J, Novel costimulators in the immune gene therapy of cancerCancer Gene Ther; 1996 3:202–213PubMedGoogle Scholar
  43. 43.
    Larchian WA, Horiguchi Y, Nair SK, Fair WR, Heston WDW, Gilboa E, Effectivenes of combined interleukin 2 and B7.1 vaccination strategy is dependent on the sequence and order: a liposome-mediated gene therapy treatment for bladder cancerClin Cancer Res;2000 6:2913–2920PubMedGoogle Scholar
  44. 44.
    Emtage PCR, Wan Y, Bramson JL, Graham FL, Gauldie J, A double recombinant adenovirus expressing the costimulatory molecule B7-1 (murine) and human IL-2 induces complete tumor regression in a murine breast adenocarcinoma modelJ Immunol 1998; 160:2531–2538PubMedGoogle Scholar
  45. 45.
    Sobol RE, Fakhrai H, Shawler D, et al. Interleukin-2 gene therapy in a patient with glioblastomaGene Ther;1995 2:164–167PubMedGoogle Scholar
  46. 46.
    Rini BI, Selk LM, Vogelzang NJ, Phase 1 study of direct intralesional gene transfer of HLA-B7 into metastatic renal carcinoma lesionsClin Cancer Res 1999; 5:2766–2772PubMedGoogle Scholar
  47. 47.
    Stewart AK, Lassam NJ, Quirt IC, et al. Adenoviral-mediated gene delivery of IL-2 in metastatic breast cancer and melanoma: results of a phase 1 clinical trialGene Ther;1999 6:350–363CrossRefPubMedGoogle Scholar
  48. 48.
    Stopeck AT, Jones A, Hersh EM, et al. Phase II study of direct intralesional gene transfer of allovectin-7, an HLA-B7/beta-2 microglobulin DNA-liposome complex, in patients with metastatic melanomaClin Cancer Res 2001; 7:2285–2291PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Ian F. Parney
    • 1
    • 6
  • Lung-Ji Chang
    • 2
  • Maxine A. Farr-Jones
    • 3
  • Chunhai Hao
    • 4
  • Michael Smylie
    • 5
  • Kenneth C. Petruk
    • 3
  1. 1.Department of Clinical Neurosciences, Southern Alberta Cancer Research Institute, and Hotchkiss Brain InstituteUniversity of CalgaryCalgaryCanada
  2. 2.Department of Molecular Genetics and Microbiology, Powell Gene Therapy Center, and McKnight Brain InstituteUniversity of FloridaGainesvilleUSA
  3. 3.Division of Neurosurgery, Department of SurgeryUniversity of Alberta EdmontonCanada
  4. 4.Department of PathologyEmory UniversityAtlantaUSA
  5. 5.Department of OncologyUniversity of AlbertaEdmontonCanada
  6. 6.Division of Neurosurgery, Department of Clinical NeurosciencesFoothills Medical CentreCalgaryCanada

Personalised recommendations