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Transfer of the murine interleukin-12 gene in vivo by a Semliki Forest virus vector induces B16 tumor regression through inhibition of tumor blood vessel formation monitored by Doppler ultrasonography

Gene Therapy volume 6, pages 606–615 (1999)Cite this article

Abstract

To elucidate further the potential of a Semliki Forest virus (SFV) vector in vivo for gene therapy, we constructed a vector, SFV-IL12, to transfer murine IL-12 genes into tumors. A single intratumoral injection of established B16 murine melanoma with SFV-IL12 resulted in a significant inhibition of tumor growth, while injection with SFV-LacZ had no effect. This antitumoral activity correlated with an increase of IFNγ production, MIG and IP-10 mRNA expression, both at the tumor site and at the periphery. In contrast, no increase in CTL- or NK cell-mediated cytotoxic response could be detected, ruling out the involvement of T and NK cell cytotoxicity. To determine how the transfer of IL-12 genes induced tumor regression, the antiangiogenicactivity of SFV-IL12 was investigated using Doppler ultrasonography (DUS). SFV-IL12 inhibited in situ neovascularization within the tumor, without affecting the resistance index of pre-existing intratumoral blood flows. In addition, histological analysis of SFV-IL12-treated tumors showed massive tumor necrosis induced by SFV-IL12 treatment. These data indicate that SFV-IL12 inhibits tumor growth through its antiangiogenic activity, demonstrated for the first time in vivo by DUS, and suggest that the SFV vector may be a novel valuable tool in tumor gene transfer.

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References

  1. Colombo MP, Forni G . Cytokine gene transfer in tumor inhibition and tumor therapy: where are we now? Immunol Today 1994 15: 48–51

    Article  CAS  PubMed  Google Scholar 

  2. Trinchieri G . Interleukin-12: a cytokine produced by antigen-presenting cells with immunoregulatory functions in the generation of T-helper cells type 1 and cytotoxic lymphocytes (review) Blood 1994 84: 4008–4027

    CAS  PubMed  Google Scholar 

  3. Chouaib S et al. Interleukin-12 induces the differentiation of major histocompatibility complex class I-primed cytotoxic T-lymphocyte precursors into allospecific cytotoxic effectors Proc Natl Acad Sci USA 1994 91: 12659–12663

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Chua AO, Chizzonite R, Desai B . Expression cloning of a human IL-12 receptor component: a new member of the cytokine receptor superfamily with strong homology with gp130 J Immunol 1994 153: 128–136

    CAS  PubMed  Google Scholar 

  5. Desai BB et al. The IL-12 receptor. II. Distribution and regulation of receptor expression J Immunol 1992 148: 3125–3132

    CAS  PubMed  Google Scholar 

  6. Manetti R et al. Natural killer cell stimulatory factor (interleukin-12 [IL-12]) induces (Th1)-type specific immune responses and inhibits the development of IL-4 producing Th2 cells J Exp Med 1993 177: 1199–1204

    Article  CAS  PubMed  Google Scholar 

  7. Chan SH et al. Induction of IFN gamma production by NK cell stimulatory factor (NKSF): characterization of the responder cells and synergy with other inducers J Exp Med 1991 173: 869–879

    Article  CAS  PubMed  Google Scholar 

  8. Gately MK et al. Administration of recombinant IL-12 to normal mice enhances cytolytic lymphocyte activity and induces production of IFNg in vivo Int Immunol 1994 6: 157–167

    Article  CAS  PubMed  Google Scholar 

  9. Brunda MJ et al. Antitumor and antimetastatic activity of interleukin-12 against murine tumors J Exp Med 1993 178: 1223–1230

    Article  CAS  PubMed  Google Scholar 

  10. Bramson JL et al. Direct intratumoral injection of an adenovirus-expressing interleukin-12 induces regression and long-lasting immunity that is associated with highly localized expression of interleukin-12 Hum Gene Ther 1996 7: 1995–2002

    Article  CAS  PubMed  Google Scholar 

  11. Caruso M et al. Adenovirus-mediated interleukin-12 gene therapy for metastatic colon carcinoma Proc Natl Acad Sci USA 1996 93: 11302–11306

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Tahara H et al. Effective eradication of established murine tumors with interleukin-12 (IL-12) gene therapy using a polycistronic retroviral vector J Immunol 1995 154: 6466–6474

    CAS  PubMed  Google Scholar 

  13. Colombo MP et al. Amount of interleukin-12 available at the tumor site is critical for tumor regression Cancer Res 1996 56: 2531–2534

    CAS  PubMed  Google Scholar 

  14. Tsung K et al. IL-12 induces T helper 1-directed antitumor response J Immunol 1997 158: 3359–3365

    CAS  PubMed  Google Scholar 

  15. Liljeström P, Garoff H . A new generation of animal cell expression vectors based on the Semliki Forest virus replicon Bio/Technology 1991 9: 1356–1361

    Article  Google Scholar 

  16. Liljestrom P . Alphavirus expression systems Curr Opin Biotechnol 1994 5: 495–500

    Article  CAS  PubMed  Google Scholar 

  17. Mossman SP et al. Protection against lethal Simian immunodeficiency virus SIVsmmPBj14 disease by a recombinant Semliki Forest virus gp160 vaccine and by a gp120 subunit vaccine J Virol 1996 70: 1953–1960

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Zhou X et al. Self-replicating Semliki Forest virus RNA as recombinant vaccine Vaccine 1994 12: 1510–1514

    Article  CAS  PubMed  Google Scholar 

  19. Zhang J et al. Cloning of human IL-12 p40 and p35 DNA into the Semliki Forest virus vector: expression of IL-12 in human tumor cells Gene Therapy 1997 4: 367–374

    Article  CAS  PubMed  Google Scholar 

  20. Folkman J . What is the evidence that tumors are angiogenesis-dependent? J Natl Cancer Inst 1990 82: 4–6

    Article  CAS  PubMed  Google Scholar 

  21. O'Reilly MS et al. Angiostatin induces and sustains dormancy of human primary tumors in mice Nature Med 1996 2: 689–692

    Article  CAS  PubMed  Google Scholar 

  22. Holmgren L, O'Reilly MS, Folkman J . Dormancy of micrometastases: balanced proliferation and apoptosis in the presence of angiogenesis suppression Nature Med 1995 1: 149–153

    Article  CAS  PubMed  Google Scholar 

  23. Millauer B et al. Glioblastoma growth inhibited in vivo by a dominant-negative Flk-1 mutant Nature 1994 367: 576–579

    Article  CAS  PubMed  Google Scholar 

  24. Hori A et al. Suppression of solid tumor growth by immunoneutralizing monoclonal antibody against human basic fibroblast growth factor Cancer Res 1991 51: 6180–6184

    CAS  PubMed  Google Scholar 

  25. Rak JW, Croix BDS, Kerbel RS . Consequences of angiogenesis for tumor progression, metastasis and cancer therapy Anticancer Drugs 1995 6: 3–18

    Article  CAS  PubMed  Google Scholar 

  26. Voest EE et al. Inhibition of angiogenesis in vivo by interleukin-12 J Natl Cancer Inst 1995 87: 581–586

    Article  CAS  PubMed  Google Scholar 

  27. Bosari S et al. Microvessel quantitation and prognosis in invasive breast carcinoma Hum Pathol 1992 23: 755–761

    Article  CAS  PubMed  Google Scholar 

  28. Cosgrove DO et al. Breast diseases: color Doppler US in differential diagnosis Radiology 1993 189: 99–104

    Article  CAS  PubMed  Google Scholar 

  29. Lassau N et al. Value of high-frequency US for preoperative assessment of skin tumors Radiographics 1997 17: 1559–1565

    Article  CAS  PubMed  Google Scholar 

  30. Lamer S et al. Radiologic assessment of intranodal vascularity in head and neck squamous cell carcinoma. Correlation with histologic vascular density Invest Radiol 1996 31: 673–679

    Article  CAS  PubMed  Google Scholar 

  31. Karlan BY, Platt LD . The current status of ultrasound and color Doppler imaging in screening for ovarian cancer Gynecol Oncol 1994 55: S28–S33

    Article  CAS  PubMed  Google Scholar 

  32. Lee WJ et al. Breast cancer angiogenesis: a quantitative morphologic and Doppler imaging study Ann Surg Oncol 1995 2: 246–251

    Article  CAS  PubMed  Google Scholar 

  33. Lassau N et al. Value of high frequency ultrasound and color Doppler ulstrasound for preoperative assessment of melanoma. American Roetgenology Society. 98th Annual Scientific Meeting Am J Roentgenol 1998 170: 197 (Abstr. 412)

    Google Scholar 

  34. Watanabe M et al. Regulation of local host-mediated anti-tumor mechanisms by cytokines: direct and indirect effects on leukocyte recruitment and angiogenesis Am J Pathol 1997 150: 1869–1880

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Kurjak A et al. An attempt to screen asymptomatic women for ovarian and endometrial cancer with transvaginal color and pulsed Doppler sonography J Ultrasound Med 1994 13: 295–301

    Article  CAS  PubMed  Google Scholar 

  36. Mu J et al. Administration of recombinant interleukin-12 prevents outgrowth of tumor cells metastasizing spontaneously to lung and lymph nodes Cancer Res 1995 55: 4404–4408

    CAS  PubMed  Google Scholar 

  37. Rakhmilevich AL et al. Gene gun-mediated skin transfection with interleukin-12 gene results in regression of established primary and metastatic murine tumors Proc Natl Acad Sci USA 1996 93: 6291–6296

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Yang Y et al. Cellular and humoral immune responses to viral antigens create barriers to lung-directed gene therapy with recombinant adenoviruses J Virol 1995 69: 2004–2015

    CAS  PubMed  PubMed Central  Google Scholar 

  39. Zhou X et al. Generation of cytotoxic and humoral immune responses using non-replicative recombinant Semliki Forest virus Proc Natl Acad Sci USA 1995 92: 3009–3013

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Nastala CL et al. Recombinant interleukin-12 administration induces tumor regression in association with interferon-gamma and nitric oxide production J Immunol 1994 153: 1697–1706

    CAS  PubMed  Google Scholar 

  41. Rodolfo M et al. Immunotherapy of experimental metastases by vaccination with interleukin gene-transduced adenocarcinoma cells sharing tumor-associated antigens. Comparison between IL-12 and IL-2 gene-transduced tumor cell vaccines J Immunol 1996 157: 5536–5542

    CAS  PubMed  Google Scholar 

  42. Barth RJJ et al. Interferon gamma and tumor necrosis factor have a role in tumor regressions mediated by murine CD8+ tumor-infiltrating lymphocytes J Exp Med 1991 173: 647–658

    Article  CAS  PubMed  Google Scholar 

  43. Schijns VE et al. IFN-gamma receptor-deficient mice generate antiviral Th1-characteristic cytokine profiles but altered antibody responses J Immunol 1994 153: 2029–2037

    CAS  PubMed  Google Scholar 

  44. Boehm U et al. Cellular responses to interferon-gamma Annu Rev Immunol 1997 15: 749–795

    Article  CAS  PubMed  Google Scholar 

  45. Emoto M et al. Differences in the angiogenesis of benign and malignant ovarian tumors, demonstrated by analyses of color Doppler ultrasound, immunohistochemistry, and microvessel density Cancer 1997 80: 899–907

    Article  CAS  PubMed  Google Scholar 

  46. Teruya-Feldstein J et al. The role of Mig, the monokine induced by interferon-gamma, and IP-10, the interferon-gamma-inducible protein-10, in tissue necrosis and vascular damage associated with Epstein–Barr virus-positive lymphoproliferative disease Blood 1997 90: 4099–4105

    CAS  PubMed  Google Scholar 

  47. Fallarino F et al. Antigen-specific regression of established tumors induced by active immunization with irradiated IL-12- but not B7–1-transfected tumor cells Int Immunol 1997 9: 1259–1269

    Article  CAS  PubMed  Google Scholar 

  48. Berglund P et al. Semliki Forest virus expression system: production of conditionally infectious recombinant particles Biotechnology (NY) 1993 11: 916–920

    CAS  Google Scholar 

  49. Asselin-Paturel C et al. Failure of TGFbeta1 and IL-12 to regulate human FasL and mTNF alloreactive cytotoxic pathways Tiss Antigen 1998 51: 242–249

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. U487 INSERM, Institut Gustave-Roussy, Villejuif, France

    C Asselin-Paturel, N Lassau, F Gay, F Mami-Chouaib & S Chouaib

  2. Département d’Imagerie Médicale, Institut Gustave-Roussy, Villejuif, France

    N Lassau & J Leclere

  3. Département d’Anatomo-Pathologie, Institut Gustave-Roussy, Villejuif, France

    J-M Guinebretière

  4. Département de Biologie Moleculaire, Université Libre de Bruxelles, Belgium

    J Zhang, F Bex & S Hallez

  5. CNRS ESA 70.57, Université de Jussieu, Paris, France

    P Peronneau

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Asselin-Paturel, C., Lassau, N., Guinebretière, JM. et al. Transfer of the murine interleukin-12 gene in vivo by a Semliki Forest virus vector induces B16 tumor regression through inhibition of tumor blood vessel formation monitored by Doppler ultrasonography. Gene Ther 6, 606–615 (1999). https://doi.org/10.1038/sj.gt.3300841

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