Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Immune response induced by retrovirus-mediated HSV-tk/GCV pharmacogene therapy in patients with glioblastoma multiforme

Gene Therapy volume 7, pages 1853–1858 (2000)Cite this article

Abstract

This study was conducted to investigate immunological components of somatic gene therapy for primary glioblastoma multiforme (GBM) in adults. It involved 13 patients treated by surgical resection of tumor with subsequent radiation therapy. Seven of them received additional herpes simplex virus thymidine kinase/ganciclovir (HSV-tk/GCV) gene therapy by direct intracerebral injection of retrovirus (RV) vector producing cells (VPC) during tumor surgery and subsequent systemic administration of GCV. Peripheral blood for FACS immunophenotyping, isolation of peripheral mononuclear cells (PMNC), and serum ELISA assays for IL-12 and soluble Fas ligand (sFasL) was collected daily during the first 4 post-operative weeks. Tumor specimens were obtained at primary or recurrent surgery and at autopsy. Tumors from gene therapy patients showed varying degrees of peritumoral necrosis around the former tumor resection cavity. Numbers of tumor-infiltrating lymphocytes found weeks after gene therapy were not significantly increased compared with primary tumors. Mitotic tumor cells were sparse close to the VPC injection sites, but abundant in brain areas somewhat distant from these sites. Serum ELISA revealed significantly increased sFasL and IL-12 levels in the gene therapy group compared with controls. Immunophenotyping of PMNC did not show a significant activation of T cells or NK cells during gene therapy. Interferon gamma secretion was evaluated by ELISPOT assays employing PMNC cocultivated with autologous tumor cells. It demonstrated an antitumor immune response in the gene therapy group, but not in the control group. These findings support the concept of in vivo induction of a systemic immune response by local intracerebral HSV-tk/GCV pharmacogene therapy for primary human GBM.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Get just this article for as long as you need it

$39.95

Learn more

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4

References

  1. Culver KW et al. In vivo gene transfer with retroviral vector-producer cells for treatment of experimental brain tumors Science 1992 256: 1550–1552

    Article  CAS  PubMed  Google Scholar 

  2. Oldfield EH et al. Gene therapy for the treatment of brain tumors using intra-tumoral transduction with the thymidine kinase gene and intravenous ganciclovir Hum Gene Ther 1993 4: 39–69

    Article  CAS  PubMed  Google Scholar 

  3. Moolten FL . Tumor chemosensitivity conferred by inserted herpes thymidine kinase genes: paradigm for a prospective cancer control strategy Cancer Res 1986 46: 5276–5281

    CAS  PubMed  Google Scholar 

  4. Danos O, Mulligan RC . Safe and efficient generation of recombinant retroviruses with amphotropic and ecotropic host ranges Proc Natl Acad Sci USA 1988 85: 6460–6464

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Izquierdo M et al. Human malignant brain tumor response to herpes simplex thymidine kinase (HSVtk)/ganciclovir gene therapy Gene Therapy 1996 3: 491–495

    CAS  PubMed  Google Scholar 

  6. Klatzmann D et al. A phase I/II study of herpes simplex virus type 1 thymidine kinase ‘suicide’ gene therapy for recurrent glioblastoma Hum Gene Ther 1998 9: 2595–2604

    CAS  PubMed  Google Scholar 

  7. Palu G et al. Gene therapy of glioblastoma multiforme via combined expression of suicide and cytokine genes: a pilot study in humans Gene Therapy 1999 6: 330–337

    Article  CAS  PubMed  Google Scholar 

  8. Ram Z et al. Therapy of malignant brain tumors by intratumoral implantation of retroviral vector-producing cells Nature Med 1997 3: 1354–1361

    Article  CAS  PubMed  Google Scholar 

  9. Shand N et al. A phase 1–2 clinical trial of gene therapy for recurrent glioblastoma multiforme by tumor transduction with the herpes simplex thymidine kinase gene followed by ganciclovir Hum Gene Ther 1999 10: 2325–2335

    Article  CAS  PubMed  Google Scholar 

  10. Puumalainen AM, Vapalahti M, Yla-Herttuala S . Gene therapy for malignant glioma patients Adv Exp Med Biol 1998 451: 505–509

    Article  CAS  PubMed  Google Scholar 

  11. Rainov NG, on behalf of the GLI328 International Study Group . a phase III clinical evaluation of herpes simplex virus type 1 thymidine kinase and ganciclovir gene therapy for previously untreated glioblastoma multiforme in adults Hum Gene Ther 2000 (in press

  12. Raffel C et al. Gene therapy for the treatment of recurrent pediatric malignant astrocytomas with in vivo transduction with the herpes simplex thymidine kinase gene/ganciclovir system Hum Gene Ther 1994 5: 863–890

    Article  CAS  PubMed  Google Scholar 

  13. Gagandeep S et al. Prodrug-activated gene therapy: involvement of an immunological component in the ‘bystander effect’ Cancer Gene Ther 1996 3: 83–88

    CAS  PubMed  Google Scholar 

  14. Freeman SM et al. In vivo studies on the mechanism of the bystander effect Cancer Gene Ther 1994 1: 326–332

    Google Scholar 

  15. Barba D, Hardin J, Sadelain M, Gage FH . Development of anti-tumor immunity following thymidine kinase-mediated killing of experimental brain tumors Proc Natl Acad Sci USA 1994 91: 4348–4352

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Hall SJ, Sanford MA, Atkinson G, Chen SH . Induction of potent antitumor natural killer cell activity by herpes simplex virus-thymidine kinase and ganciclovir therapy in an orthotopic mouse model of prostate cancer Cancer Res 1998 58: 3221–3225

    CAS  PubMed  Google Scholar 

  17. Mullen CA et al. Ganciclovir chemoablation of herpes thymidine kinase suicide gene-modified tumors produces tumor necrosis and induces systemic immune responses Hum Gene Ther 1998 9: 2019–2030

    Article  CAS  PubMed  Google Scholar 

  18. Vile RG et al. Generation of an anti-tumor immune response in a non-immunogenic tumor: HSVtk killing in vivo stimulates a mononuclear cell infiltrate and a Th1-like profile of intratumoral cytokine expression Int J Cancer 1997 71: 365–369

    Article  Google Scholar 

  19. Yamamoto S et al. Herpes simplex virus thymidine kinase/ganciclovir-mediated killing of tumor cell induces tumor-specific cytotoxic T cells in mice Cancer Gene Ther 1997 4: 91–96

    PubMed  Google Scholar 

  20. Andreansky S et al. Treatment of intracranial gliomas inimmunocompetent mice using herpes simplex viruses thatexpress murine interleukins Gene Therapy 1998 5: 121–130

    Article  CAS  PubMed  Google Scholar 

  21. Weber FW et al. MRI characteristics and histological changes in glioblastoma after gene therapy Front Radiat Ther Oncol 1999 33: 244–252

    Article  CAS  PubMed  Google Scholar 

  22. Nakayama Y et al. Serum levels of interleukin-12 in patients with gastrointestinal cancer Anticancer Res 2000 20: 635–640

    CAS  PubMed  Google Scholar 

  23. Lissoni P et al. Physiopathology of IL-12 in human solid neoplasms: blood levels of IL-12 in early or advanced cancer patients, and their variations with surgery and immunotherapy J Biol Regul Homeost Agents 1998 12: 38–41

    CAS  PubMed  Google Scholar 

  24. Lissoni P et al. Increased blood concentrations of interleukin-12 are associated with a longer survival in untreatable metastatic solid tumor patients: preliminary observations Int J Biol Markers 1997 12: 125–127

    Article  CAS  PubMed  Google Scholar 

  25. Toyozaki T et al. Levels of soluble Fas ligand in myocarditis Am J Cardiol 1998 82: 246–248

    Article  CAS  PubMed  Google Scholar 

  26. Tanaka M, Suda T, Takahashi T, Nagata S . Expression of the functional soluble form of human fas ligand in activated lymphocytes EMBO J 1995 14: 1129–1135

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Rogulski KR et al. Pronounced antitumor effects and tumor radiosensitization of double suicide gene therapy Clin Cancer Res 1997 3: 2081–2088

    CAS  PubMed  Google Scholar 

  28. Vrionis FD et al. Preservation of the bystander cytocidal effect of irradiated herpes simplex virus thymidine kinase (HSV-tk) modified tumor cells J Neurooncol 1996 30: 225–236

    Article  CAS  PubMed  Google Scholar 

  29. Kramm CM et al. Systemic activation of the immune system during ganciclovir treatment following intratumoral grafting of HSVtk retroviral vector producer cells in an adolescent ependymoma patient Cancer Gene Ther 2000 (in press

  30. Kirk AD et al. The human antiporcine cellular repertoire. In vitro studies of acquired and innate cellular responsiveness Transplantation 1993 55: 924–931

    Article  CAS  PubMed  Google Scholar 

  31. Körholz D et al. The role of interleukin-10 (IL-10) in IL-15-mediated T-cell responses Blood 1997 90: 4513–4521

    PubMed  Google Scholar 

  32. Long Z et al. Molecular evaluation of biopsy and autopsy specimens from patients receiving in vivo retroviral gene therapy Hum Gene Ther 1999 10: 733–740

    Article  CAS  PubMed  Google Scholar 

  33. Rodenburg RJ, van den Hoogen FH, van de Putte LB, van Venrooij WJ . Peripheral blood monocytes of rheumatoid arthritis patients do not express elevated TNF alpha IL-1beta and IL-8 mRNA levels. A comparison of monocyte isolation procedures J Immunol Methods 1998 221: 169–175

    Article  CAS  PubMed  Google Scholar 

  34. Butterfield LH et al. Generation of human T-cell responses to an HLA-A21-restricted peptide epitope derived from alpha-fetoprotein Cancer Res 1994 59: 3134–3142

    Google Scholar 

  35. Larsson M et al. A recombinant vaccinia virus based ELISPOT assay detects high frequencies of Pol-specific CD8 T cells in HIV-1-positive individuals AIDS 1999 13: 767–777

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This study was supported in part by grants 015VE1997 and 2794A/0087H from the Federal State of Saxony-Anhalt (NGR), grant 0311661/1402 (BioRegio) from the Federal Ministry of Education and Research of Germany (NGR, CMK), grant KR1711/1–1 from the German Research Council, DFG (CMK), grant SFB 503-C6 from the German Research Council, DFG (UB), grants from the Elterninitiative Kinderkrebsklinik Duesseldorf eV (CMK, UB, DK), and by Novartis Pharma GmbH, Nuremberg, Germany.

Author information

Authors and Affiliations

  1. Department of Neurosurgery, Martin-Luther-University Halle-Wittenberg, Halle, Germany

    N G Rainov, V Heidecke & W Burkert

  2. Department of Pediatric Hematology and Oncology, University Children's Hospital, Heinrich-Heine-University, Duesseldorf, Germany

    C M Kramm, U Banning & D Körholz

  3. Institute of Medical Immunology, Martin-Luther-University, Germany

    D Riemann

  4. Institute of Pathology, Martin-Luther-University Halle, Germany

    H-J Holzhausen

  5. Novartis Pharma GmbH, Nuremberg, Germany

    K J Burger

Authors
  1. N G Rainov
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C M Kramm
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. U Banning
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D Riemann
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. H-J Holzhausen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. V Heidecke
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. K J Burger
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. W Burkert
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. D Körholz
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Rainov, N., Kramm, C., Banning, U. et al. Immune response induced by retrovirus-mediated HSV-tk/GCV pharmacogene therapy in patients with glioblastoma multiforme. Gene Ther 7, 1853–1858 (2000). https://doi.org/10.1038/sj.gt.3301311

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301311

Keywords

  • ganciclovir
  • gene therapy
  • glioblastoma
  • peripheral blood mononuclear cells
  • retrovirus
  • thymidine kinase

This article is cited by

Search

Advanced search

Quick links