Skip to main content
SpringerLink
Log in

Inhibition of established subcutaneous and metastatic murine tumors by intramuscular electroporation of the interleukin-12 gene

  • Original Paper
  • Published:
Journal of Biomedical Science

Abstract

In vivo electroporation (EP) of the murine interleukin-12 (IL-12) gene in an expression plasmid (pIL-12) was evaluated for antitumor activity. EP transfer of pIL-12 into mouse quadriceps muscles elicited significant levels of serum IL-12 and interferon-γ. Intramuscular EP of pIL-12 resulted in complete regression or substantial inhibition of 38C13 B-cell lymphoma, whereas pIL-12 delivered by gene gun or intramuscular injection without EP showed little therapeutic effect. Impressive antitumor activity by intramuscular EP was also demonstrated in animals with advanced malignant disease. At day 14 after 38C13 tumor inoculation, all animals were found to carry large tumors and to have metastases; without treatment, most died within a week. A single intramuscular EP of pIL-12 resulted in regression of 50% of large subcutaneous tumors and significantly prolonged the lifespan of these animals. Moreover, animals that were previously cured of 38C13 tumors by in vivo EP treatment significantly suppressed tumor growth when challenged 60 days later. In vivo EP of the IL-12 gene was also effective in suppressing subcutaneous and lung metastatic tumors of CT-26 colon adenocarcinoma and B16F1 melanoma cells. Together, these results show that intramuscular electrotransfer of the IL-12 gene may represent a simple and effective strategy for cancer treatment.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Aihara H, Miyazaki J. Gene transfer into muscle by electroporation in vivo. Nat Biotechnol 16:867–870;1998.

    Google Scholar 

  2. Bergman Y, Haimovich J. Characterization of a carcinogen-induced murine B lymphocyte cell line of C3H/eB origin. Eur J Immunol 7:413–417;1977.

    Google Scholar 

  3. Boggio K, Nicoletti G, Di Carlo E, Cavallo F, Landuzzi L, Melani C, Giovarelli M, Rossi I, Nanni P, De Giovanni C, Bouchard P, Wolf S, Modesti A, Musiani P, Lollini PL, Colombo MP, Forni G. Interleukin 12-mediated prevention of spontaneous mammary adenocarcinomas in two lines of Her-2/neu transgenic mice. J Exp Med 188:589–596;1998.

    Google Scholar 

  4. Bramson JL, Hitt M, Addison CL, Muller WJ, Gauldie J, Graham FL. 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 7:1995–2002;1996.

    Google Scholar 

  5. Brunda MJ, Luistro L, Warrier RR, Wright RB, Hubbard BR, Murphy M, Wolf SF, Gately MK. Antitumor and antimetastatic activity of interleukin 12 against murine tumors. J Exp Med 178:1223–1230;1993.

    Google Scholar 

  6. Campbell MJ, Esserman L, Byars NE, Allison AC, Levy R. Idiotype vaccination against murine B cell lymphoma. Humoral and cellular requirements for the full expression of antitumor immunity. J Immunol 145:1029–1036;1990.

    Google Scholar 

  7. Campbell MJ, Esserman L, Levy R. Immunotherapy of established murine B cell lymphoma. Combination of idiotype immunization and cyclophosphamide. J Immunol 141:3227–3233;1988.

    Google Scholar 

  8. Chan SH, Perussia B, Gupta JW, Kobayashi M, Pospisil M, Young HA, Wolf SF, Young D, Clark SC, Trinchieri G. Induction of interferon gamma production by natural killer cell stimulatory factor: characterization of the responder cells and synergy with other inducers. J Exp Med 173:869–879;1991.

    Google Scholar 

  9. Chen L, Chen D, Block E, O'Donnell M, Kufe DW, Clinton SK. Eradication of murine bladder carcinoma by intratumor injection of a bicistronic adenoviral vector carrying cDNAs for the IL-12 heterodimer and its inhibition by the IL-12 p40 subunit homodimer. J Immunol 159:351–359;1997.

    Google Scholar 

  10. Chow YH, Chiang BL, Lee YL, Chi WK, Lin WC, Chen YT, Tao MH. Development of Th1 and Th2 populations and the nature of immune responses to hepatitis B virus DNA vaccines can be modulated by codelivery of various cytokine genes. J Immunol 160:1320–1329;1998.

    Google Scholar 

  11. Cohen J. IL-12 deaths: Explanation and a puzzle. Science 270:908;1995.

    Google Scholar 

  12. Corbett TH, Griswold DPJ, Roberts BJ, Peckham JC, Schabel FMJ. Tumor induction relationships in development of transplantable cancers of the colon in mice for chemotherapy assays, with a note on carcinogen structure. Cancer Res 35:2434–2439;1975.

    Google Scholar 

  13. Coughlin CM, Salhany KE, Wysocka M, Aruga E, Kurzawa H, Chang AE, Hunter CA, Fox JC, Trinchieri G, Lee WMF. Interleukin-12 and interleukin-18 synergistically induce murine tumor regression which involves inhibition of angiogenesis. J Clin Invest 101:1441–1452;1998.

    Google Scholar 

  14. Dev SB, Hofmann GA. Electrochemotherapy — a novel method of cancer treatment. Cancer Treat Rev 20:105–115;1994.

    Google Scholar 

  15. Draghia Akli R, Fiorotto ML, Hill LA, Malone PB, Deaver DR, Schwartz RJ. Myogenic expression of an injectable protease-resistant growth hormone-releasing hormone augments long-term growth in pigs. Nat Biotechnol 17:1179–1183;1999.

    Google Scholar 

  16. Gately MK, Renzetti LM, Magram J, Stern AS, Adorini L, Gubler U, Presky DH. The interleukin-12/interleukin-12-receptor system: role in normal and pathologic immune responses. Annu Rev Immunol 16:495–521;1998.

    Google Scholar 

  17. Gately MK, Warrier RR, Honasoge S, Carvajal DM, Faherty DA, Connaughton SE, Anderson TD, Sarmiento U, Hubbard BR, Murphy M. Administration of recombinant IL-12 to normal mice enhances cytolytic lymphocyte activity and induces production of IFN-gamma in vivo. Int Immunol 6:157–167;1994.

    Google Scholar 

  18. Germann T, Gately MK, Schoenhaut DS, Lohoff M, Mattner F, Fischer S, Jin SC, Schmitt E, Rude E. Interleukin-12/T cell stimulating factor, a cytokine with multiple effects on T helper type 1 (Th1) but not on Th2 cells. Eur J Immunol 23:1762–1770;1993.

    Google Scholar 

  19. Goto T, Nishi T, Tamura T, Dev SB, Takeshima H, Kochi M, Yoshizato K, Kuratsu J, Sakata T, Hofmann GA, Ushio Y. Highly efficient electro-gene therapy of solid tumor by using an expression plasmid for the herpes simplex virus thymidine kinase gene. Proc Natl Acad Sci U S A 97:354–359;2000.

    Google Scholar 

  20. Heller R, Jaroszeski M, Atkin A, Moradpour D, Gilbert R, Wands J, Nicolau C. In vivo gene electroinjection and expression in rat liver. FEBS Lett 389:225–228;1996.

    Google Scholar 

  21. Hyacinthe M, Jaroszeski MJ, Dang VV, Coppola D, Karl RC, Gilbert RA, Heller R. Electrically enhanced drug delivery for the treatment of soft tissue sarcoma. Cancer 85:409–417;1999.

    Google Scholar 

  22. Kozarsky KF, Wilson JM. Gene therapy: Adenovirus vectors. Curr Opin Genet Dev 3:499–503;1993.

    Google Scholar 

  23. Kwak LW, Young HA, Pennington RW, Weeks SD. Vaccination with syngeneic, lymphoma-derived immunoglobulin idiotype combined with granulocyte-macrophage colony-stimulating factor primes mice for a protective T-cell response. Proc Natl Acad Sci USA 93:10972–10977;1996.

    Google Scholar 

  24. Ledley FD. Nonviral gene therapy: the promise of genes as pharmaceutical products. Hum Gene Ther 6:1129–1144;1995.

    Google Scholar 

  25. Lee YL, Tao MH, Chow YH, Chiang BL. Construction of vectors expressing bioactive heterodimeric and single-chain murine interleukin-12 for gene therapy. Hum Gene Ther 9:457–465;1998.

    Google Scholar 

  26. Lode HN, Reisfeld RA. Targeted cytokines for cancer immunotherapy. Immunol Res 21:279–288;2000.

    Google Scholar 

  27. Lohr F, Lo DY, Zaharoff DA, Hu K, Zhang X, Li Y, Zhao Y, Dewhirst MW, Yuan F, Li CY. Effective tumor therapy with plasmid-encoded cytokines combined with in vivo electroporation. Cancer Res 61:3281–3284;2001.

    Google Scholar 

  28. Maruyama H, Sugawa M, Moriguchi Y, Imazeki I, Ishikawa Y, Ataka K, Hasegawa S, Ito Y, Higuchi N, Kazama JJ, Gejyo F, Miyazaki JI. Continuous erythropoietin delivery by muscle-targeted gene transfer using in vivo electroporation. Hum Gene Ther 11:429–437;2000.

    Google Scholar 

  29. Meko JB, Yim JH, Tsung K, Norton JA. High cytokine production and effective antitumor activity of a recombinant vaccinia virus encoding murine interleukin 12. Cancer Res 55:4765–4770;1995.

    Google Scholar 

  30. Mu J, Zou JP, Yamamoto N, Tsutsui T, Tai XG, Kobayashi M, Herrmann S, Fujiwara H, Hamaoka T. Administration of recombinant interleukin 12 prevents outgrowth of tumor cells metastasizing spontaneously to lung and lymph nodes. Cancer Res 55:4404–4408;1995.

    Google Scholar 

  31. Mulligan RC. The basic science of gene therapy. Science 260:926–932;1993.

    Google Scholar 

  32. Muramatsu T, Mizutani Y, Ohmori Y, Okumura J. Comparison of three nonviral transfection methods for foreign gene expression in early chicken embryos in ovo. Biochem Biophys Res Commun 230:376–380;1997.

    Google Scholar 

  33. Nanda GS, Sun FX, Hofmann GA, Hoffman RM, Dev SB. Electroporation enhances therapeutic efficacy of anticancer drugs: treatment of human pancreatic tumor in animal model. Anticancer Res 18:1361–1366;1998.

    Google Scholar 

  34. Nastala CL, Edington HD, McKinney TG, Tahara H, Nalesnik MA, Brunda MJ, Gately MK, Wolf SF, Schreiber RD, Storkus WJ, et al. Recombinant IL-12 administration induces tumor regression in association with IFN-gamma production. J Immunol 153:1697–1706;1994.

    Google Scholar 

  35. Neumann E, Schaefer Ridder M, Wang Y, Hofschneider PH. Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J 1:841–845;1982.

    Google Scholar 

  36. Nishi T, Yoshizato K, Yamashiro S, Takeshima H, Sato K, Hamada K, Kitamura I, Yoshimura T, Saya H, Kuratsu J, Ushio Y. High-efficiency in vivo gene transfer using intraarterial plasmid DNA injection following in vivo electroporation. Cancer Res 56:1050–1055;1996.

    Google Scholar 

  37. Nishioka Y, Hirao M, Robbins PD, Lotze MT, Tahara H. Induction of systemic and therapeutic antitumor immunity using intratumoral injection of dendritic cells genetically modified to express interleukin 12. Cancer Res 59:4035–4041;1999.

    Google Scholar 

  38. Orange JS, Salazar Mather TP, Opal SM, Spencer RL, Miller AH, McEwen BS, Biron CA. Mechanism of interleukin 12-mediated toxicities during experimental viral infections: role of tumor necrosis factor and glucocorticoids. J Exp Med 181:901–914;1995.

    Google Scholar 

  39. Oshima Y, Sakamoto T, Yamanaka I, Nishi T, Ishibashi T, Inomata H. Targeted gene transfer to corneal endothelium in vivo by electric pulse. Gene Ther 5:1347–1354;1998.

    Google Scholar 

  40. Peng LS, Penichet ML, Morrison SL. A single-chain IL-12 IgG3 antibody fusion protein retains antibody specificity and IL-12 bioactivity and demonstrates antitumor activity. J Immunol 163:250–258;1999.

    Google Scholar 

  41. Rakhmilevich AL, Turner J, Ford MJ, McCabe D, Sun WH, Sondel PM, Grota K, Yang NS. Gene gun-mediated skin transfection with interleukin 12 gene results in regression of established primary and metastatic murine tumors. Proc Natl Acad Sci USA 93:6291–6296;1996.

    Google Scholar 

  42. Rizzuto G, Cappelletti M, Mennuni C, Wiznerowicz M, Demartis A, Maione D, Ciliberto G, Monica NL, Fattori E. Gene electrotransfer results in a high-level transduction of rat skeletal muscle and corrects anemia of renal failure. Hum Gene Ther 11:1891–1900;2000.

    Google Scholar 

  43. Rols MP, Delteil C, Golzio M, Dumond P, Cros S, Teissie J. In vivo electrically mediated protein and gene transfer in murine melanoma. Nat Biotechnol 16:168–171;1998.

    Google Scholar 

  44. Tahara H, Lotze MT. Antitumor effects of interleukin-12 (IL-12): applications for the immunotherapy and gene therapy of cancer. Gene Ther 2:96–106;1995.

    Google Scholar 

  45. Tahara H, Zeh H Jr, Storkus WJ, Poppo I, Watkins SC, Gubler U, Wolf SF, Robbins PD, Lotze MT. Fibroblasts genetically engineered to secrete interleukin 12 can suppress tumor growth and induce antitumor immunity to a murine melanoma in vivo. Cancer Res 54:182–189;1994.

    Google Scholar 

  46. Tahara H, Zitvogel L, Storkus WJ, Zeh HJ, McKinney TG, Schreiber RD, Gubler U, Robbins PD, Lotze MT. Effective eradication of established murine tumors with IL-12 gene therapy using a polycistronic retroviral vector. J Immunol 154:6466–6474;1995.

    Google Scholar 

  47. Talmadge JE, Fidler IJ. Cancer metastasis is selective or random depending on the patent tumor population. Nature 297:593–594;1982.

    Google Scholar 

  48. Titomirov AV, Sukharev S, Kistanova E. In vivo electroporation and stable transformation of skin cells of newborn mice by plasmid DNA. Biochim Biophys Acta 1088:131–134;1991.

    Google Scholar 

  49. Trinchieri G, Scott P. Interleukin-12: Basic principles and clinical applications. Curr Top Microbiol Immunol 238:57–78;1999.

    Google Scholar 

  50. Wolff JA, Ludtke JJ, Acsadi G, Williams P, Jani A. Long-term persistence of plasmid DNA and foreign gene expression in mouse muscle. Hum Mol Genet 1:363–369;1992.

    Google Scholar 

  51. Wolff JA, Malone RW, Williams P, Chong W, Acsadi G, Jani A, Felgner PL. Direct gene transfer into mouse muscle in vivo. Science 247:1465–1468;1990.

    Google Scholar 

  52. Yamashita YI, Shimada M, Hasegawa H, Minagawa R, Rikimaru T, Hamatsu T, Tanaka S, Shirabe K, Miyazaki JI, Sugimachi K. Electroporation-mediated interleukin-12 gene therapy for hepatocellular carcinoma in the mice model. Cancer Res 61:1005–1012;2001.

    Google Scholar 

  53. Zhou P, Sieve MC, Bennett J, Kwon-Chung KJ, Tewari RP, Gazzinelli RT, Sher A, Seder RA. IL-12 prevents mortality in mice infected with Histoplasma capsulatum through induction of IFN-gamma. J Immunol 155:785–795;1995.

    Google Scholar 

  54. Zimmermann U. Electric field-mediated fusion and related electrical phenomena. Biochim Biophys Acta 694:227–277;1982.

    Google Scholar 

  55. Zitvogel L, Tahara H, Cai Q, Storkus WJ, Muller G, Wolf SF, Gately MK, Robbins PD, Lotze MT. Construction and characterization of retroviral vectors expressing biologically active human interleukin-12. Hum Gene Ther 5:1493–1506;1994.

    Google Scholar 

  56. Zitvogel L, Tahara H, Robbins PD, Storkus WJ, Clarke MR, Nalesnik MA, Lotze MT. Cancer immunotherapy of established tumors with IL-12. Effective delivery by genetically engineered fibroblasts. J Immunol 155:1393–1403;1995.

    Google Scholar 

  57. Zou JP, Yamamoto N, Fujii T, Takenaka H, Kobayashi M, Herrmann SH, Wolf SF, Fujiwara H, Hamaoka T. Systemic administration of rIL-12 induces complete tumor regression and protective immunity: response is correlated with a striking reversal of suppressed IFN-gamma production by anti-tumor T cells. Int Immunol 7:1135–1145;1995.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. National Health Research Institutes, Taipei

    Shan-Chih Lee & Cheng-Wen Wu

  2. Institute of Biomedical Sciences Academia Sinica, 128 Yen-Jiou Yuan Rd., Sec. 2 Nankang, 11529, Taipei, Taiwan (ROC)

    Shan-Chih Lee, Chang-Jer Wu, Pin-Yi Wu, Yi-Ling Huang & Mi-Hua Tao PhD

Authors
  1. Shan-Chih Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Chang-Jer Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Pin-Yi Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yi-Ling Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Cheng-Wen Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mi-Hua Tao PhD
    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

Lee, SC., Wu, CJ., Wu, PY. et al. Inhibition of established subcutaneous and metastatic murine tumors by intramuscular electroporation of the interleukin-12 gene. J Biomed Sci 10, 73–86 (2003). https://doi.org/10.1007/BF02256000

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02256000

Key words

Search

Navigation