Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1998 Mar 15;101(6):1441–1452. doi: 10.1172/JCI1555

Interleukin-12 and interleukin-18 synergistically induce murine tumor regression which involves inhibition of angiogenesis.

C M Coughlin 1, K E Salhany 1, M Wysocka 1, E Aruga 1, H Kurzawa 1, A E Chang 1, C A Hunter 1, J C Fox 1, G Trinchieri 1, W M Lee 1
PMCID: PMC508700  PMID: 9502787

Abstract

The antitumor effect and mechanisms activated by murine IL-12 and IL-18, cytokines that induce IFN-gamma production, were studied using engineered SCK murine mammary carcinoma cells. In syngeneic A/J mice, SCK cells expressing mIL-12 or mIL-18 were less tumorigenic and formed tumors more slowly than control cells. Neither SCK.12 nor SCK.18 cells protected significantly against tumorigenesis by distant SCK cells. However, inoculation of the two cell types together synergistically protected 70% of mice from concurrently injected distant SCK cells and 30% of mice from SCK cells established 3 d earlier. Antibody neutralization studies revealed that the antitumor effects of secreted mIL-12 and mIL-18 required IFN-gamma. Interestingly, half the survivors of SCK.12 and/or SCK.18 cells developed protective immunity suggesting that anti-SCK immunity is unlikely to be responsible for protection. Instead, angiogenesis inhibition, assayed by Matrigel implants, appeared to be a property of both SCK.12 and SCK.18 cells and the two cell types together produced significantly greater systemic inhibition of angiogenesis. This suggests that inhibition of tumor angiogenesis is an important part of the systemic antitumor effect produced by mIL-12 and mIL-18.

Full Text

The Full Text of this article is available as a PDF (705.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abrams J. S., Roncarolo M. G., Yssel H., Andersson U., Gleich G. J., Silver J. E. Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples. Immunol Rev. 1992 Jun;127:5–24. doi: 10.1111/j.1600-065x.1992.tb01406.x. [DOI] [PubMed] [Google Scholar]
  2. Angiolillo A. L., Sgadari C., Taub D. D., Liao F., Farber J. M., Maheshwari S., Kleinman H. K., Reaman G. H., Tosato G. Human interferon-inducible protein 10 is a potent inhibitor of angiogenesis in vivo. J Exp Med. 1995 Jul 1;182(1):155–162. doi: 10.1084/jem.182.1.155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baskar S., Ostrand-Rosenberg S., Nabavi N., Nadler L. M., Freeman G. J., Glimcher L. H. Constitutive expression of B7 restores immunogenicity of tumor cells expressing truncated major histocompatibility complex class II molecules. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5687–5690. doi: 10.1073/pnas.90.12.5687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bloom E. T., Horvath J. A. Cellular and molecular mechanisms of the IL-12-induced increase in allospecific murine cytolytic T cell activity. Implications for the age-related decline in CTL. J Immunol. 1994 May 1;152(9):4242–4254. [PubMed] [Google Scholar]
  5. Boehm U., Klamp T., Groot M., Howard J. C. Cellular responses to interferon-gamma. Annu Rev Immunol. 1997;15:749–795. doi: 10.1146/annurev.immunol.15.1.749. [DOI] [PubMed] [Google Scholar]
  6. Car B. D., Eng V. M., Schnyder B., LeHir M., Shakhov A. N., Woerly G., Huang S., Aguet M., Anderson T. D., Ryffel B. Role of interferon-gamma in interleukin 12-induced pathology in mice. Am J Pathol. 1995 Dec;147(6):1693–1707. [PMC free article] [PubMed] [Google Scholar]
  7. Cavallo F., Signorelli P., Giovarelli M., Musiani P., Modesti A., Brunda M. J., Colombo M. P., Forni G. Antitumor efficacy of adenocarcinoma cells engineered to produce interleukin 12 (IL-12) or other cytokines compared with exogenous IL-12. J Natl Cancer Inst. 1997 Jul 16;89(14):1049–1058. doi: 10.1093/jnci/89.14.1049. [DOI] [PubMed] [Google Scholar]
  8. Colombo M. P., Vagliani M., Spreafico F., Parenza M., Chiodoni C., Melani C., Stoppacciaro A. Amount of interleukin 12 available at the tumor site is critical for tumor regression. Cancer Res. 1996 Jun 1;56(11):2531–2534. [PubMed] [Google Scholar]
  9. Coughlin C. M., Wysocka M., Kurzawa H. L., Lee W. M., Trinchieri G., Eck S. L. B7-1 and interleukin 12 synergistically induce effective antitumor immunity. Cancer Res. 1995 Nov 1;55(21):4980–4987. [PubMed] [Google Scholar]
  10. Coughlin C. M., Wysocka M., Trinchieri G., Lee W. M. The effect of interleukin 12 desensitization on the antitumor efficacy of recombinant interleukin 12. Cancer Res. 1997 Jun 15;57(12):2460–2467. [PubMed] [Google Scholar]
  11. Hanahan D., Folkman J. Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis. Cell. 1996 Aug 9;86(3):353–364. doi: 10.1016/s0092-8674(00)80108-7. [DOI] [PubMed] [Google Scholar]
  12. Hsieh C. S., Macatonia S. E., Tripp C. S., Wolf S. F., O'Garra A., Murphy K. M. Development of TH1 CD4+ T cells through IL-12 produced by Listeria-induced macrophages. Science. 1993 Apr 23;260(5107):547–549. doi: 10.1126/science.8097338. [DOI] [PubMed] [Google Scholar]
  13. Kleinman H. K., McGarvey M. L., Liotta L. A., Robey P. G., Tryggvason K., Martin G. R. Isolation and characterization of type IV procollagen, laminin, and heparan sulfate proteoglycan from the EHS sarcoma. Biochemistry. 1982 Nov 23;21(24):6188–6193. doi: 10.1021/bi00267a025. [DOI] [PubMed] [Google Scholar]
  14. Majewski S., Marczak M., Szmurlo A., Jablonska S., Bollag W. Interleukin-12 inhibits angiogenesis induced by human tumor cell lines in vivo. J Invest Dermatol. 1996 May;106(5):1114–1118. doi: 10.1111/1523-1747.ep12340161. [DOI] [PubMed] [Google Scholar]
  15. Martinotti A., Stoppacciaro A., Vagliani M., Melani C., Spreafico F., Wysocka M., Parmiani G., Trinchieri G., Colombo M. P. CD4 T cells inhibit in vivo the CD8-mediated immune response against murine colon carcinoma cells transduced with interleukin-12 genes. Eur J Immunol. 1995 Jan;25(1):137–146. doi: 10.1002/eji.1830250124. [DOI] [PubMed] [Google Scholar]
  16. Micallef M. J., Ohtsuki T., Kohno K., Tanabe F., Ushio S., Namba M., Tanimoto T., Torigoe K., Fujii M., Ikeda M. Interferon-gamma-inducing factor enhances T helper 1 cytokine production by stimulated human T cells: synergism with interleukin-12 for interferon-gamma production. Eur J Immunol. 1996 Jul;26(7):1647–1651. doi: 10.1002/eji.1830260736. [DOI] [PubMed] [Google Scholar]
  17. Micallef M. J., Yoshida K., Kawai S., Hanaya T., Kohno K., Arai S., Tanimoto T., Torigoe K., Fujii M., Ikeda M. In vivo antitumor effects of murine interferon-gamma-inducing factor/interleukin-18 in mice bearing syngeneic Meth A sarcoma malignant ascites. Cancer Immunol Immunother. 1997 Jan;43(6):361–367. doi: 10.1007/s002620050345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nastala C. L., Edington H. D., McKinney T. G., Tahara H., Nalesnik M. A., Brunda M. J., Gately M. K., Wolf S. F., Schreiber R. D., Storkus W. J. Recombinant IL-12 administration induces tumor regression in association with IFN-gamma production. J Immunol. 1994 Aug 15;153(4):1697–1706. [PubMed] [Google Scholar]
  19. O'Reilly M. S., Boehm T., Shing Y., Fukai N., Vasios G., Lane W. S., Flynn E., Birkhead J. R., Olsen B. R., Folkman J. Endostatin: an endogenous inhibitor of angiogenesis and tumor growth. Cell. 1997 Jan 24;88(2):277–285. doi: 10.1016/s0092-8674(00)81848-6. [DOI] [PubMed] [Google Scholar]
  20. O'Reilly M. S., Holmgren L., Shing Y., Chen C., Rosenthal R. A., Moses M., Lane W. S., Cao Y., Sage E. H., Folkman J. Angiostatin: a novel angiogenesis inhibitor that mediates the suppression of metastases by a Lewis lung carcinoma. Cell. 1994 Oct 21;79(2):315–328. doi: 10.1016/0092-8674(94)90200-3. [DOI] [PubMed] [Google Scholar]
  21. Okamura H., Tsutsi H., Komatsu T., Yutsudo M., Hakura A., Tanimoto T., Torigoe K., Okura T., Nukada Y., Hattori K. Cloning of a new cytokine that induces IFN-gamma production by T cells. Nature. 1995 Nov 2;378(6552):88–91. doi: 10.1038/378088a0. [DOI] [PubMed] [Google Scholar]
  22. Passaniti A., Taylor R. M., Pili R., Guo Y., Long P. V., Haney J. A., Pauly R. R., Grant D. S., Martin G. R. A simple, quantitative method for assessing angiogenesis and antiangiogenic agents using reconstituted basement membrane, heparin, and fibroblast growth factor. Lab Invest. 1992 Oct;67(4):519–528. [PubMed] [Google Scholar]
  23. Sgadari C., Angiolillo A. L., Tosato G. Inhibition of angiogenesis by interleukin-12 is mediated by the interferon-inducible protein 10. Blood. 1996 May 1;87(9):3877–3882. [PubMed] [Google Scholar]
  24. Sgadari C., Farber J. M., Angiolillo A. L., Liao F., Teruya-Feldstein J., Burd P. R., Yao L., Gupta G., Kanegane C., Tosato G. Mig, the monokine induced by interferon-gamma, promotes tumor necrosis in vivo. Blood. 1997 Apr 15;89(8):2635–2643. [PubMed] [Google Scholar]
  25. Song C. W., Kang M. S., Rhee J. G., Levitt S. H. Vascular damage and delayed cell death in tumours after hyperthermia. Br J Cancer. 1980 Feb;41(2):309–312. doi: 10.1038/bjc.1980.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tahara H., Zeh H. J., 3rd, Storkus W. J., Pappo I., Watkins S. C., Gubler U., Wolf S. F., Robbins P. D., Lotze M. T. Fibroblasts genetically engineered to secrete interleukin 12 can suppress tumor growth and induce antitumor immunity to a murine melanoma in vivo. Cancer Res. 1994 Jan 1;54(1):182–189. [PubMed] [Google Scholar]
  27. Tahara H., Zitvogel L., Storkus W. J., Zeh H. J., 3rd, McKinney T. G., Schreiber R. D., Gubler U., Robbins P. D., Lotze M. T. Effective eradication of established murine tumors with IL-12 gene therapy using a polycistronic retroviral vector. J Immunol. 1995 Jun 15;154(12):6466–6474. [PubMed] [Google Scholar]
  28. Tan J., Newton C. A., Djeu J. Y., Gutsch D. E., Chang A. E., Yang N. S., Klein T. W., Hua Y. Injection of complementary DNA encoding interleukin-12 inhibits tumor establishment at a distant site in a murine renal carcinoma model. Cancer Res. 1996 Aug 1;56(15):3399–3403. [PubMed] [Google Scholar]
  29. Trinchieri G. Interleukin-12: a proinflammatory cytokine with immunoregulatory functions that bridge innate resistance and antigen-specific adaptive immunity. Annu Rev Immunol. 1995;13:251–276. doi: 10.1146/annurev.iy.13.040195.001343. [DOI] [PubMed] [Google Scholar]
  30. Tsung K., Meko J. B., Peplinski G. R., Tsung Y. L., Norton J. A. IL-12 induces T helper 1-directed antitumor response. J Immunol. 1997 Apr 1;158(7):3359–3365. [PubMed] [Google Scholar]
  31. Tsutsui H., Nakanishi K., Matsui K., Higashino K., Okamura H., Miyazawa Y., Kaneda K. IFN-gamma-inducing factor up-regulates Fas ligand-mediated cytotoxic activity of murine natural killer cell clones. J Immunol. 1996 Nov 1;157(9):3967–3973. [PubMed] [Google Scholar]
  32. Ushio S., Namba M., Okura T., Hattori K., Nukada Y., Akita K., Tanabe F., Konishi K., Micallef M., Fujii M. Cloning of the cDNA for human IFN-gamma-inducing factor, expression in Escherichia coli, and studies on the biologic activities of the protein. J Immunol. 1996 Jun 1;156(11):4274–4279. [PubMed] [Google Scholar]
  33. Voest E. E., Kenyon B. M., O'Reilly M. S., Truitt G., D'Amato R. J., Folkman J. Inhibition of angiogenesis in vivo by interleukin 12. J Natl Cancer Inst. 1995 Apr 19;87(8):581–586. doi: 10.1093/jnci/87.8.581. [DOI] [PubMed] [Google Scholar]
  34. Watanabe M., McCormick K. L., Volker K., Ortaldo J. R., Wigginton J. M., Brunda M. J., Wiltrout R. H., Fogler W. E. Regulation of local host-mediated anti-tumor mechanisms by cytokines: direct and indirect effects on leukocyte recruitment and angiogenesis. Am J Pathol. 1997 May;150(5):1869–1880. [PMC free article] [PubMed] [Google Scholar]
  35. Wysocka M., Kubin M., Vieira L. Q., Ozmen L., Garotta G., Scott P., Trinchieri G. Interleukin-12 is required for interferon-gamma production and lethality in lipopolysaccharide-induced shock in mice. Eur J Immunol. 1995 Mar;25(3):672–676. doi: 10.1002/eji.1830250307. [DOI] [PubMed] [Google Scholar]
  36. Xie Q. W., Whisnant R., Nathan C. Promoter of the mouse gene encoding calcium-independent nitric oxide synthase confers inducibility by interferon gamma and bacterial lipopolysaccharide. J Exp Med. 1993 Jun 1;177(6):1779–1784. doi: 10.1084/jem.177.6.1779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Yoshimoto T., Okamura H., Tagawa Y. I., Iwakura Y., Nakanishi K. Interleukin 18 together with interleukin 12 inhibits IgE production by induction of interferon-gamma production from activated B cells. Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3948–3953. doi: 10.1073/pnas.94.8.3948. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Yu W. G., Yamamoto N., Takenaka H., Mu J., Tai X. G., Zou J. P., Ogawa M., Tsutsui T., Wijesuriya R., Yoshida R. Molecular mechanisms underlying IFN-gamma-mediated tumor growth inhibition induced during tumor immunotherapy with rIL-12. Int Immunol. 1996 Jun;8(6):855–865. doi: 10.1093/intimm/8.6.855. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES