Cancer Immunology, Immunotherapy

, Volume 61, Issue 10, pp 1671–1682 | Cite as

Intratumoral electroporation of IL-12 cDNA eradicates established melanomas by Trp2180–188-specific CD8+ CTLs in a perforin/granzyme-mediated and IFN-γ-dependent manner: application of Trp2180–188 peptides

  • Jeong-Im Sin
  • Jae-Bok Park
  • In Hee Lee
  • Daehan Park
  • Youn Seok Choi
  • Jongseon Choe
  • Esteban Celis
Original article

Abstract

Intratumoral electroporation (IT-EP) with IL-12 cDNA (IT-EP/IL12) can lead to the eradication of established B16 melanoma tumors in mice. Here, we explore the immunological mechanism of the antitumor effects generated by this therapy. The results show that IT-EP/IL12 applied only once resulted in eradication in 70% animals with large established B16 tumors. Tumor eradication required the participation of CD8+ T cells, but not CD4+ T cells and NK cells. IT-EP/IL12 induced antigen-specific CD8+ T cell responses against the immunodominant Trp2180–188 epitope and generated a systemic response, resulting in significant therapeutic effects against distal, untreated tumors. The therapeutic effect of IT-EP/IL12 was absent in perforin-deficient mice, indicating that tumor elimination occurred through conventional perforin/granzyme lysis by CTLs. Moreover, this therapy induced some degree of immunological memory that protected approximately one-third of the cured mice against a subsequent tumor challenge. Moreover, antitumor efficacy and long-term protection against B16 were significantly improved by concurrent Trp2 peptide immunization through more induction of Ag-specific CTL responses and more attraction of IFN-γ-expressing CD8+ T cells into tumor sites. The antitumor effect of IT-EP/IL12 required the participation of IFN-γ, which was shown to induce MHC class I expression on B16 cells and increase the lytic activity of the CD8+ CTL generated by IT-EP/IL12. The results from these animal studies may help in the development of IT-EP/IL12 for cancer patients.

Keywords

Antitumor immunity Electroporation IL-12 Melanoma 

References

  1. 1.
    Lohr F, Lo DY, Zaharoff DA, Hu K, Zhang X, Li Y, Zhao Y, Dewhirst MW, Yuan F, Li CY (2001) Effective tumor therapy with plasmid-encoded cytokines combined with in vivo electroporation. Cancer Res 61(8):3281–3284PubMedGoogle Scholar
  2. 2.
    Robertson MJ, Soiffer RJ, Wolf SF, Manley TJ, Donahue C, Young D, Hermann SH, Ritz J (1992) Responses of human natural killer (NK) cells to NK cell stimulatory factor (NKSF): cytolytic activity and proliferation of NK cells are differentially regulated by NKSF. J Exp Med 175:779–788PubMedCrossRefGoogle Scholar
  3. 3.
    Kobayashi M, Fitz L, Ryan M, Hewick RM, Clark SC, Chan S, Loudon R, Sherman F, Perussia B, Trinchieri G (1989) Identification and purification of natural killer cell stimulator factor (NKSF): a cytokine with multiple biologic effects on human lymphocytes. J Exp Med 170:827–845PubMedCrossRefGoogle Scholar
  4. 4.
    Germann T, Gately MK, Schoenhaut DS, Lohoff M, Mattner F, Fischer S, Jin SC, Schmitt E, Rude E (1993) 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 23:1762–1770PubMedCrossRefGoogle Scholar
  5. 5.
    Coughlin CM, Salhany KE, Gee MS, LaTemple DC, Kotenko S, Ma X, Gri G, Wysocka M, Kim JE, Liu L, Liao F, Farber JM, Pestka S, Trinchieri G, Lee WM (1998) Tumor cell responses to IFNgamma affect tumorigenicity and response to IL-12 therapy and antiangiogenesis. Immunity 9(1):25–34PubMedCrossRefGoogle Scholar
  6. 6.
    Gately MK, Warrier RR, Honasoge S, Carvajal DM, Faherty DA, Connaughton SE, Anderson TD, Sarmiento U, Hubbard BR, Murphy M (1994) Administration of recombinant IL-12 to normal mice enhances cytolytic lymphocyte activity and induces production of IFN-gamma in vivo. Int Immunol 6(1):157–167PubMedCrossRefGoogle Scholar
  7. 7.
    Lucas ML, Heller L, Coppola D, Heller R (2002) IL-12 plasmid delivery by in vivo electroporation for the successful treatment of established subcutaneous B16.F10 melanoma. Mol Ther 5(6):668–675PubMedCrossRefGoogle Scholar
  8. 8.
    Li S, Zhang L, Torrero M, Cannon M, Barret R (2005) Administration route- and immune cell activation-dependent tumor eradication by IL12 electrotransfer. Mol Ther 12:942–949PubMedCrossRefGoogle Scholar
  9. 9.
    Yamashita Y, Shimada M, Hasegawa H, Minagawa R, Rikimaru T, Hamatsu T, Tanaka S, Shirabe K, Miyazaki JI, Sugimachi K (2001) Electroporation-mediated interleukin-12 gene therapy for hepatocellular carcinoma in the mice model. Cancer Res 61(3):1005–1012PubMedGoogle Scholar
  10. 10.
    Cemazar M, Jarm T, Sersa G (2010) Cancer electrogene therapy with interleukin-12. Curr Gene Ther 10(4):300–311PubMedCrossRefGoogle Scholar
  11. 11.
    Daud AI, DeConti RC, Andrews S, Urbas P, Riker AI, Sondak VK, Munster PN, Sullivan DM, Ugen KE, Messina JL, Heller R (2008) Phase I trial of interleukin-12 plasmid electroporation in patients with metastatic melanoma. J Clin Oncol 26:5896–5903PubMedCrossRefGoogle Scholar
  12. 12.
    Pavlin D, Cemazar M, Cör A, Sersa G, Pogacnik A (2011) Tozon N (2011) Electrogene therapy with interleukin-12 in canine mast cell tumors. Radiol Oncol 45:31–39PubMedCrossRefGoogle Scholar
  13. 13.
    Bae SH, Park YJ, Choi YS, Park JB, Kim MS, Sin JI (2007) Therapeutic synergy of human papillomavirus E7 subunit vaccines plus cisplatin in an animal tumor model: causal involvement of increased sensitivity of cisplatin-treated tumors to CTL-mediated killing in therapeutic synergy. Clin Cancer Res 13(1):341–349PubMedCrossRefGoogle Scholar
  14. 14.
    Ye GW, Park JB, Park YJ, Choi YS, Sin JI (2007) Increased sensitivity of radiated murine cervical cancer tumors to E7 subunit vaccine-driven CTL-mediated killing induces synergistic antitumor activity. Mol Ther 15(8):1564–1570PubMedCrossRefGoogle Scholar
  15. 15.
    Sin JI, Kim JM, Bae SH, Lee IH, Park JS, Ryoo HM (2009) Adoptive transfer of human papillomavirus E7-specific CTL enhances tumor chemoresponse through the Perforin/Granzyme-mediated Pathway. Mol Ther 17(5):906–913PubMedCrossRefGoogle Scholar
  16. 16.
    Sin JI, Kim JJ, Arnold RL, Shroff KE, McCallus D, Pachuk C, McElhiney SP, Wolf MW, Pompa-de Bruin SJ, Higgins TJ, Ciccarelli RB, Weiner DB (1999) IL-12 gene as a DNA vaccine adjuvant in a herpes mouse model: IL-12 enhances Th1 type CD4+ T cell mediated protective immunity against herpes simplex virus-2 challenge. J Immunol 162:2912–2921PubMedGoogle Scholar
  17. 17.
    Sin JI (2009) Suppression of antitumor protective cytotoxic T lymphocyte responses to a human papillomavirus 16 E7 DNA vaccine by coinjection of interleukin-12 cDNA: involvement of nitric oxide in immune suppression. Immunology 128:e707–e717PubMedCrossRefGoogle Scholar
  18. 18.
    Kim TY, Myoung HJ, Kim JH, Moon IS, Kim TG, Ahn WS, Sin JI (2002) Both E7 and CpG-ODN are required for protective immunity against challenge with human papillomavirus 16 (E6/E7)-immortalized tumor cells: involvement of CD4+ and CD8+ T cells in protection. Cancer Res 62:7234–7240PubMedGoogle Scholar
  19. 19.
    Corr M, von Damm A, Lee DJ, Tighe H (1999) In vivo priming by DNA injection occurs predominantly by antigen transfer. J Immunol 163:4721–4727PubMedGoogle Scholar
  20. 20.
    Sin JI, Hong SH, Park YJ, Park JB, Choi YS, Kim MS (2006) Antitumor therapeutic effects of E7 subunit and DNA vaccines in an animal cervical cancer model: antitumor efficacy of E7 therapeutic vaccines is dependent on tumor sizes, vaccine doses, and vaccine delivery routes. DNA Cell Biol 25(5):277–286PubMedCrossRefGoogle Scholar
  21. 21.
    Robbins PF, Sette A, Appella E, Rosenberg SA (1995) Recognition of multiple epitopes in the human melanoma antigen gp100 by tumor-infiltrating T lymphocytes associated with in vivo tumor regression. J Immunol 154:3961–3968PubMedGoogle Scholar
  22. 22.
    Wang RF, Appella E, Kawakami Y, Kang X, Rosenberg SA (1996) Identification of TRP-2 as a human tumor antigen recognized by cytotoxic T lymphocytes. J Exp Med 184:2207–2216PubMedCrossRefGoogle Scholar
  23. 23.
    Bloom MB, Perry-Lalley D, Robbins PF, Li Y, el-Gamil M, Rosenberg SA, Yang JC (1997) Identification of tyrosinase-related protein 2 as a tumor rejection antigen for the B16 melanoma. J Exp Med 185(3):453–459PubMedCrossRefGoogle Scholar
  24. 24.
    Finkelstein SE, Heimann DM, Klebanoff CA, Antony PA, Gattinoni L, Hinrichs CS, Hwang LN, Palmer DC, Spiess PJ, Surman DR, Wrzesiniski C, Yu Z, Rosenberg SA, Restifo NP (2004) Bedside to bench and back again: how animal models are guiding the development of new immunotherapies for cancer. J Leukoc Biol 76(2):333–337PubMedCrossRefGoogle Scholar
  25. 25.
    Guevara-Patiño JA, Engelhorn ME, Turk MJ, Liu C, Duan F, Rizzuto G, Cohen AD, Merghoub T, Wolchok JD, Houghton AN (2006) Optimization of a self antigen for presentation of multiple epitopes in cancer immunity. J Clin Invest 116(5):1382–1390PubMedCrossRefGoogle Scholar
  26. 26.
    Amos S, Pegram HJ, Westwood JA, John LB, Devaud C, Clarke CJ, Restifo NP, Smyth MJ, Darcy PK, Kershaw MH (2011) Adoptive immunotherapy combined with intratumoral TLR agonist delivery eradicates established melanoma in mice. Cancer Immunol Immunother 60(5):671–683PubMedCrossRefGoogle Scholar
  27. 27.
    Cho H, Lee YR, Celis E (2011) Interferon gamma limits the effectiveness of melanoma peptide vaccines. Blood 117(1):135–144PubMedCrossRefGoogle Scholar
  28. 28.
    Kishida T, Asada H, Satoh E, Tanaka S, Shinya M, Hirai H, Iwai M, Tahara H, Imanishi J, Mazda O (2001) In vivo electroporation-mediated transfer of interleukin-12 and interleukin-18 genes induces significant antitumor effects against melanoma in mice. Gene Ther 8(16):1234–1240PubMedCrossRefGoogle Scholar
  29. 29.
    Tamura T, Nishi T, Goto T, Takeshima H, Dev SB, Ushio Y, Sakata T (2001) Intratumoral delivery of interleukin 12 expression plasmids with in vivo electroporation is effective for colon and renal cancer. Hum Gene Ther 12(10):1265–1276PubMedCrossRefGoogle Scholar
  30. 30.
    Shibata M, Ito Y, Morimoto J, Kusakabe K, Yoshinaka R, Otsuki Y (2006) In vivo electrogene transfer of interleukin-12 inhibits tumor growth and lymph node and lung metastases in mouse mammary carcinomas. J Gene Med 8(3):335–352PubMedCrossRefGoogle Scholar
  31. 31.
    Pavlin D, Cemazar M, Kamensek U, Tozon N, Pogacnik A, Sersa G (2009) Local and systemic antitumor effect of intratumoral and peritumoral IL-12 electrogene therapy on murine sarcoma. Cancer Biol Ther 8(22):2114–2122PubMedCrossRefGoogle Scholar
  32. 32.
    Heller L, Cruz YL, Ferraro B, Yang H, Heller R (2010) Plasmid injection and application of electric pulses alter endogenous mRNA and protein expression in B16.F10 mouse melanomas. Cancer Gene Ther 17(12):864–871PubMedCrossRefGoogle Scholar
  33. 33.
    Ferraro B, Heller LC, Cruz YL, Guo S, Donate A, Heller R (2011) Evaluation of delivery conditions for cutaneous plasmid electrotransfer using a multielectrode array. Gene Ther 18(5):496–500PubMedCrossRefGoogle Scholar
  34. 34.
    Sauter B, Albert ML, Francisco L, Larsson M, Somersan S, Bhardwaj N (2000) Consequences of cell death: exposure to necrotic tumor cells, but not primary tissue cells or apoptotic cells, induces the maturation of immunostimulatory dendritic cells. J Exp Med 191(3):423–434PubMedCrossRefGoogle Scholar
  35. 35.
    Leonard JP, Sherman ML, Fisher GL, Buchanan LJ, Larsen G, Atkins MB, Sosman JA, Dutcher JP, Vogelzang NJ, Ryan JL (1997) Effects of single-dose interleukin-12 exposure on interleukin-12-associated toxicity and interferon-gamma production. Blood 90:2541–2548PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Jeong-Im Sin
    • 1
  • Jae-Bok Park
    • 2
  • In Hee Lee
    • 3
  • Daehan Park
    • 4
  • Youn Seok Choi
    • 5
  • Jongseon Choe
    • 1
  • Esteban Celis
    • 6
  1. 1.Department of Microbiology, School of MedicineKangwon National UniversityChuncheonKorea
  2. 2.Department of Pathology, School of MedicineCatholic University of DaeguNamguKorea
  3. 3.Department of Nephrology, School of MedicineCatholic University of DaeguNamguKorea
  4. 4.Department of Plastic Surgery, School of MedicineCatholic University of DaeguNamguKorea
  5. 5.Department of Obstetrics and Gynecology, School of MedicineCatholic University of DaeguNamguKorea
  6. 6.Immunology Program, Department of Oncologic Sciences, Moffitt Cancer CenterUniversity of South FloridaTampaUSA

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