Cancer Immunology, Immunotherapy

, Volume 31, Issue 5, pp 305–311 | Cite as

Active specific immunotherapy with vaccinia colon oncolysate enhances the immunomodulatory and antitumor effects of interleukin-2 and interferon α in a murine hepatic metastasis model

  • Pedro J. Arroyo
  • Jerry A. Bash
  • Marc K. Wallack
Original articles


The role of cytokines as primary or adjuvant antineoplastic agents has been well established. Interleukin-2 (IL-2) and the interferons have, particularly, proven to be effective antitumor agents when given alone, and seem to act synergistically on the eradication of metastases from immunogenic tumors. Active specific immunotherapy, in the form of viral oncolysates, has also shown effectiveness in cancer therapy. Bearing this in mind, we decided to combine these agents in an adjuvant triple regimen and compare their effectiveness to other treatments in terms of tumor burden and survival in a murine colon cancer hepatic metastases model. BALB/c mice were injected with CC-36, a weakly immunogenic murine colon adenocarcinoma, intrasplenically, to produce artificial liver metastases. The animals were divided into one control group and seven treatment groups receiving either vaccinia colon oncolysate (VCO), IL-2, interferon-α (IFNα) alone, or combinations of these agents. Half the animals were followed for survival and the other half were sacrificed at the end of the experiment for quantification of tumor burden. The blood of the sacrificed animals was utilized in a series of immunological tests in order to demonstrate the cytolytic potential of the peripheral blood lymphocytes (PBL) in each treatment group, as well as to characterize phenotypically the cells acting as effectors. The tripleadjuvant regimen group was by far the most effective treatment group, demonstrating 100% survival and a significant reduction in tumor burden when compared to other groups. Furthermore, the PBL from the animals in this group showed 69.4% lysis of the CC-36 target cells in vitro. These effector lymphocytes were characterized as ASMG1-/Lyt2.2+ cytolytic lymphocytes. We conclude that these lymphocytes were stimulated by the administration of VCO and further augmented by the immunomodulation of the cytokines given in the triple regimen, and that such a regimen might prove beneficial in the treatment of established hepatic metastases from weakly immunogenic tumors.


Peripheral Blood Lymphocyte Hepatic Metastasis Vaccinia Cytolytic Activity Nodule Count 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Ansel S, Blangy D (1984) In vivo induction of tumor-specific immunity by glycolipid extracts of SV40-transformed cells. Int J Cancer 34: 555CrossRefPubMedGoogle Scholar
  2. 2.
    Barnavon Y, Iwaki H, Bash JA, Brettschneider F, Hilsenbeck S, Darnell E, Wallack MK (1988) Treatment of murine hepatic metastases with vaccinia colon oncolysates and IL-2. J Surg Res 45: 523CrossRefPubMedGoogle Scholar
  3. 3.
    Barnavon Y, Iwaki H, Bash JA, Wallack MK (1988) Vaccinia colon oncolysates immunotherapy for murine hepatic metastases can be modulated with low-dose interleukin-2. Am Surg 54: 696PubMedGoogle Scholar
  4. 4.
    Brunda MJ, Bellantoni D, Sulich V (1987) In vivo antitumor activity of combinations of alpha interferon and interleukin-2 in a murine model. Correlation of efficacy with the induction of cytotoxic cells resembling natural killer cells. Int J Cancer 40: 365CrossRefPubMedGoogle Scholar
  5. 5.
    Cameron RB, McIntosh JK, Rosenberg SA (1988) Synergistic antitumor effects of combination immunotherapy with recombinant interleukin-2 and a recombinant hybrid alpha interferon in the treatment of established murine hepatic metastases. Cancer Res 48: 5810PubMedGoogle Scholar
  6. 6.
    Chen BD, Najor F (1987) Macrophage activation by interferon alpha and beta is associated with a loss of proliferative capacity: role of interferon alpha and beta in the regulation of macrophage proliferation and function. Cell Immunol 106: 343CrossRefPubMedGoogle Scholar
  7. 7.
    Corbett TH, Griswold DP, Robert BJ, Peckman JC, Schabel FM (1975) Tumor induction relationships in development of transplantable cancers of the colon in mice for chemotherapy assays with a note on carcinogenic structure. Cancer Res 35: 2434PubMedGoogle Scholar
  8. 8.
    Fekete E (1938) A comparative morphological study of the mammary gland in high and low tumor strain in mice. Am J Pathol 14: 557PubMedPubMedCentralGoogle Scholar
  9. 9.
    Fertsch D, Schoenberg DR, Germain RN, Tou JYL, Vogel SN (1987) Induction of macrophage Ia antigen expression by rIFN-gamma and down-regulation by IFN-a/a and dexamethasone are mediated by changes in steady state levels of Ia mRNA. J Immunol 139: 244PubMedGoogle Scholar
  10. 10.
    Fidler IJ, Heicappell R, Saiki I, Grutter G, Horisberger MA, Nuesch J (1987) Direct antiproliferative effect of recombinant human interferon-alpha B/D hybrids on human tumor cell lines. Cancer Res 47: 2020PubMedGoogle Scholar
  11. 11.
    Foon KA (1989) Biological response modifiers: the new immunotherapy. Cancer Res 49: 1621PubMedGoogle Scholar
  12. 12.
    Fujiwara H, Shimizu Y, Takai Y, Wakamiya N, Ueda S, Kato S, Hamaoka T (1984) The augmentation of tumor-specific immunity by virus help. I. Demonstration of vaccinia virus reactive helper T-cell activity involved in enhanced induction of cytotoxic T-lymphocytes and antibody responses. Eur J Immunol 14: 171CrossRefPubMedGoogle Scholar
  13. 13.
    Galili N, Mavor B, Asjo B, Klein A (1976) Induction of immune responsiveness in a genetically low-responsive tumor-host combination by chemical modification of immunogen. Eur J Immunol 6: 473CrossRefPubMedGoogle Scholar
  14. 14.
    Goldstein D, Laszlo J (1986) Interferon therapy in cancer: from imaginon to interferon. Cancer Res 46: 4315PubMedGoogle Scholar
  15. 15.
    Gresser I (1977) On the varied biologic effects of interferon. Cell Immunol 34: 406CrossRefPubMedGoogle Scholar
  16. 16.
    Gutterman JU, Blumeschein GR, Alexanian R, Yap HY, Bugdai AU, Cabanillas F, Hortobagui GN, Hersh EM, Rasmussen SL, Harmon M, Kramer M, Pestka S (1980) Leukocyte interferon-induced tumor regression in human metastatic breast cancer, multiple myeloma and malignant lymphoma. Ann Intern Med 93: 399CrossRefPubMedGoogle Scholar
  17. 17.
    Herberman RB, Ortaldo JR, Timonen T, Reynolds CW, Djeu JY, Pestka S, Stanton J (1981) Interferon and natural killer (NK) cells. Texas Rep Biol Med 41: 590Google Scholar
  18. 18.
    Heron I, Hokland M, Berg K (1981) Enhanced expression of beta-2-microglobulin and HLA antigens on human lymphoid cells by interferon. Proc Natl Acad Sci USA 75: 6515Google Scholar
  19. 19.
    Hersey P, Edward A, D'Alessamdro G, MacDonald M (1986) Phase II study of vaccinia melanoma cell lysates (VMCL) as adjuvant to surgical treatment of stage II melanoma. Cancer Immunol Immunother 22: 221PubMedGoogle Scholar
  20. 20.
    Hicks NJ, Morris AG, Burke DC (1981) Partial reversion of transformed phenotype of murine sarcoma virus-transformed cells in the presence of interferon: a possible mechanism for the antitumor effect of interferon. J Cell Sci 49: 225PubMedGoogle Scholar
  21. 21.
    Iwaki H, Barnavon Y, Bash JA, Wallack MK (1989) Vaccinia virusinfected CC-36 colon tumor cell lysates stimulate cellular responses in vitro and protect syngeneic BALB/c mice from tumor cell challenge. J Surg Oncol 40: 90CrossRefPubMedGoogle Scholar
  22. 22.
    Jett JR, Mantovani A, Herberman RB (1980) Augmentation of human monocyte-mediated cytolysis by interferon. Cell Immunol 540: 425CrossRefGoogle Scholar
  23. 23.
    Kataoka T, Matsuura N, Oh-hashi F, Suhara Y (1985) Treatment regimen and host T-cell dependent therapeutic effect of interferon in mouse solid tumors. Cancer Res 45: 3548PubMedGoogle Scholar
  24. 24.
    Krigel RL, Padavic-Shaller KA, Rudolph AR, Litwin S, Konrad M, Bradley EC, Comis RL (1988) A phase I study of recombinant interleukin-2 plus recombinant beta interferon. Cancer Res 48: 3875PubMedGoogle Scholar
  25. 25.
    Kuribayashi K, Gillis S, Kern DE, Henney CS (1981) Murine NK cell cultures: effects of interleukin-2 and interferon on cell growth and cytotoxic reactivity. J Immunol 126: 2321PubMedGoogle Scholar
  26. 26.
    Lafreniere R, Rosenberg SA (1985) Adoptive immunotherapy of experimental hepatic metastases with lymphokine-activated killer cells (LAK) and recombinant interleukin-2 can mediate the regression of both immunogenic and non-immunogenic sarcomas and an adenocarcinoma in a murine system. J Immunol 135: 4273PubMedGoogle Scholar
  27. 27.
    Lafreniere R, Rosenberg SA (1986) A novel approach to the generation and identification of experimental hepatic metastases in a murine model. J Natl Cancer Inst 76: 309PubMedGoogle Scholar
  28. 28.
    Lindenman J, Klein PA (1967) Viral oncolysis increased immunogenicity of host cell antigen associated with influenza virus. J Exp Med 126: 93CrossRefGoogle Scholar
  29. 29.
    Lotze MT, Chang AE, Seipp CA, Simpson CG, Vetto JT, Rosenberg SA (1986) High-dose recombinant interleukin-2 in the treatment of patients with disseminated cancer. J Am Med Assoc 256: 3117CrossRefGoogle Scholar
  30. 30.
    Lotze MT, Matory YT, Rayner AA, Ettingehausen SE, Vetto JT, Seipp CA, Rosenberg SA (1986) Clinical effects and toxicity of interleukin-2 in patients with cancer. Cancer 58: 2764CrossRefPubMedGoogle Scholar
  31. 31.
    Martin WJ, Wunderlich JR, Fletcher F, Inman JK (1971) Enhanced immunogenicity of chemical coated syngeneic tumor cells. Proc Natl Acad Sci USA 68: 469CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Merigan TC (1988) Human interferon as a therapeutic agent. N Engl J Med 318: 1458CrossRefPubMedGoogle Scholar
  33. 33.
    Mule JJ, Shu S, Schwarz SL, Rosenberg SA (1984) Successful adoptive immunotherapy of established pulmonary metastases with LAK cells and recombinant IL-2. Science 225: 1487CrossRefPubMedGoogle Scholar
  34. 34.
    Quesada JR, Reuben JR, Manning JT (1984) Alpha interferon for induction of remission in hairy cell leukemia. N Engl J Med 310: 15CrossRefPubMedGoogle Scholar
  35. 35.
    Reed LJ, Muench H (1938) A simple method of estimating fifty percent endpoints. Am J Hyg 27: 493Google Scholar
  36. 36.
    Rehberg E, Kelder B, Hoal EG, Pestka S (1982) Specific molecular activities of recombinant and hybrid leukocyte interferons. J Biol Chem 257: 11497PubMedGoogle Scholar
  37. 37.
    Rosenberg SA, Mule JJ, Speiss PJ, Reichert CM, Schwarz SL (1985) Regression of established pulmonary metastases and subcutaneous tumor mediated by the systemic administration of high-dose recombinant interleukin-2. J Exp Med 161: 1169CrossRefPubMedGoogle Scholar
  38. 38.
    Rosenberg SA, Speiss PJ, Lafreniere R (1986) A new approach to the adoptive immunotherapy of cancer with tumor-infiltrating lymphocytes. Science 233: 1318CrossRefPubMedGoogle Scholar
  39. 39.
    Rosenberg SA, Lotze MT, Muul LM, Chang AE, Avis FP, Leitman S, Linehan WM, Robertston CN, Lee RE, Rubin JT, Seipp CA, Simpson CG, White DE (1987) A progress report on the treatment of 157 patients with advanced cancer using lymphokine-activated killer cells and interleukin-2 or high dose interleukin-2 alone. N Engl J Med 316: 889CrossRefPubMedGoogle Scholar
  40. 40.
    Rosenberg SA, Schwarz SL, Spiess PJ (1988) Combination immunotherapy for cancer: Synergistic antitumor interactions of interleukin-2, alfa interferon and tumor-infiltrating lymphocytes. J Natl Cancer Inst 80: 1393CrossRefPubMedGoogle Scholar
  41. 41.
    Sato N, Michaelides MC, Wallack MK (1981) Characterization of tumorigenicity, mortality, metastasis, and splenomegaly of two cultured murine colon lines. Cancer Res 41: 2267PubMedGoogle Scholar
  42. 42.
    Sato N, Michaelides MC, Wallack MK (1983) Transplantation immunity and cross-protection of two cultured murine colon lines. J Natl Cancer Inst 70: 231Google Scholar
  43. 43.
    Shimizu Y, Fujiwara H, Ueda S, Wakamiya N, Kato S, Hamaoka T (1984) The augmentation of tumor-specific immunity by virus help. II. Enhanced induction of tumor antigens by vaccinia virus-reactive helper T-cells. Eur J Immunol 14: 839CrossRefPubMedGoogle Scholar
  44. 44.
    Shimizu Y, Hasumi K, Masubuchi K, Okudaira Y (1988) Immunotherapy of tumor-bearing mice utilizing virus help. Cancer Immunol Immunother 27: 223CrossRefPubMedGoogle Scholar
  45. 45.
    Speiss PJ, Yang JC, Rosenberg SA (1987) Tumor infiltrating lymphocytes expanded in recombinant interleukin-2 mediate potent antitumor activity in vivo. J Natl Cancer Inst 79: 1067Google Scholar
  46. 46.
    Spiegel RJ (1987) The alpha interferons: clinical overview. Semin Oncol 14 [Suppl 2]: 1PubMedGoogle Scholar
  47. 47.
    Takeichi N, Austin FC, Tsuneyuki O, Boone CW (1978) Augmented immunogenicity produced by interferon with influenza virus as compared to Moloney sarcoma virus. Cancer Res 38: 4580PubMedGoogle Scholar
  48. 48.
    Tamura T, Sasaki Y, Shinkai T, Euuchi K, Sakurai M, Fujiwara Y, Nakagawa K, Minato K, Bungo M, Aijo N (1989) Phase I study of combination therapy with interleukin-2 and beta interferon in patients with advanced malignancy. Cancer Res 49: 730PubMedGoogle Scholar
  49. 49.
    Thompson JA, Peace DJ, Klarnet JP, Kern E, Greenberg PD, Cheever MA (1986) Eradication of disseminated murine leukemia by treatment with high-dose interleukin-2. J Immunol 137: 3675PubMedGoogle Scholar
  50. 50.
    Wallack MK (1979) Vaccinia virus augmented vaccines a new form of immunotherapy. GANN Monogr Cancer Res 23: 273Google Scholar
  51. 51.
    Wallack MK, Steplewski Z, Koprowski H, Rosato E, George J, Hullihan B, Johnson J (1977) A new approach in specific active immunotherapy. Cancer 39: 560CrossRefPubMedGoogle Scholar
  52. 52.
    Wallack MK, Meyer M, Bourgoin A, Dore JF, Leftheriotis E, Carcagne J, Koprowski H (1983) A preliminary trial of vaccinia oncolysates in the treatment of recurrent melanoma with serologic responses to treatment. J Biol Response Mod 2: 586PubMedGoogle Scholar
  53. 53.
    Wallack MK, McNally KR, Leftheriotis E, Siegler H, Balch C, Wanebo H, Bartolucci A, Bash JA (1986) A Southeastern Cancer Study Group phase I/II trial with vaccinia melanoma oncolysates. Cancer 57: 649CrossRefPubMedGoogle Scholar
  54. 54.
    Wan YJ, Orrison BM, Lieberman R, Lazarovici P, Ozato K (1987) Induction of major histocompatibility class I antigens by interferons in undifferentiated F9 cells. J Cell Physiol 130: 276CrossRefPubMedGoogle Scholar
  55. 55.
    Welsh RM (1978) Cytotoxic cells induced during lymphocytic choriomeningitis virus infection of mice. I. Characterization of natural killer cell induction. J Exp Med 148: 163CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • Pedro J. Arroyo
    • 1
  • Jerry A. Bash
    • 1
  • Marc K. Wallack
    • 1
  1. 1.Department of SurgeryMount Sinai Medical CenterMiami BeachUSA

Personalised recommendations