Abstract
We have previously shown that BALB/c-derived EMT6 mammary tumours transfected with interleukin (IL)-2 have decreased hypoxia compared to parental tumours, due to increased vascularization. Since hypoxia is a critical factor in the response of tumours to radiation treatment, we compared the radiation response of IL-2-transfected tumours to that of parental EMT6 tumours. Because the IL-2 tumours have an altered host cell composition, which could affect the interpretation of radiation sensitivity as measured by clonogenic cells, we employed flow cytometric analysis to determine the proportion of tumour cells vs host cells in each tumour type. Using this approach, we were able to correct the plating efficiency based on the number of actual tumour cells derived from tumours, making the comparison of the two types of tumours possible. We also excluded the possibility that cytotoxic T-cells present in EMT6/IL-2 tumours could influence the outcome of the clonogenic cell survival assay, by demonstrating that the plating efficiency of cells derived from EMT6/IL-2 tumours remained unchanged after depletion of Thy-1+cells. The in vivo radiation response results demonstrated that IL-2-transfected tumours were more sensitive to radiation than parental EMT6 tumours. The hypoxic fraction of the EMT6/IL-2 tumours growing in vivo was markedly decreased relative to parental EMT6 tumours thus the increased sensitivity results from the increased vascularity we have previously observed in these tumours. These results indicate the potential therapeutic benefit of combining radiation and immunotherapy in the treatment of tumours. © 2000 Cancer Research Campaign
Similar content being viewed by others
Article PDF
Change history
16 November 2011
This paper was modified 12 months after initial publication to switch to Creative Commons licence terms, as noted at publication
References
Badalament RA, Cibas ES, Reuter VE, Fair WR and Melamed MR (1987) Flow cytometric analysis of primary adenocarcinoma of the bladder. J Urol 137: 1159–1162
Braunschweiger PG, Basrur V, Santos O, Adessa A, Houdek P and Markoe AM (1996) Radioresistance in murine solid tumors induced by interleukin-1. Radiat Res 145: 150–156
Cameron RB, Spiess PJ and Rosenberg SA (1990) Synergistic antitumor activity of tumor-infiltrating lymphocytes, interleukin 2, and local tumor irradiation. Studies on the mechanism of action. J Exp Med 171: 249–263
Chadwick KH and Leenhouts HP (1973) A molecular theory of cell survival. Physics Med Biol 18: 78–87
Chang AY and Keng PC (1987) Potentiation of radiation cytotoxicity by recombinant interferons, a phenomenon associated with increased blockage at the G2-M phase of the cell cycle. Cancer Res 47: 4338–4341
Chiang CS, Syljuasen RG, Hong JH, Wallis A, Dougherty GJ and McBride WH (1997) Effects of IL-3 gene expression on tumor response to irradiation in vitro and in vivo. Cancer Res 57: 3899–3903
Colombo MP, Ferrari G, Stoppacciaro A, Parenza M, Rodolfo M, Mavilio F and Parmiani G (1991) Granulocyte colony-stimulating factor gene transfer suppresses tumorigenicity of a murine adenocarcinoma in vivo. J Exp Med 173: 889–897
Evans SM, Jenkins WT, Joiner B, Lord EM and Koch CJ (1996) 2-Nitroimidazole (EF5) binding predicts radiation resistance in individual 9L s.c. tumors. Cancer Res 56: 405–411
Fearon ER, Pardoll DM, Itaya T, Golumbek P, Levitsky HI, Simons JW, Karasuyama H, Vogelstein B and Frost P (1990) Interleukin-2 production by tumor cells bypasses T helper function in the generation of an antitumor response. Cell 60: 397–403
Gansbacher B, Zier K, Daniels B, Cronin K, Bannerji R and Gilboa E (1990) Interleukin 2 gene transfer into tumor cells abrogates tumorigenicity and induces protective immunity. J Exp Med 172: 1217–1224
Gatenby RA, Kessler HB, Rosenblum JS, Coia LR, Moldofsky PJ, Hartz WH and Broder GJ (1988) Oxygen distribution in squamous cell carcinoma metastases and its relationship to outcome of radiation therapy. Int J Radiat Oncol Biol Physics 14: 831–838
Gerber M, Guichard M, Pioch Y and Dubois JB (1989) The influence of interleukin-2, feeder cells, and timing of irradiation on the radiosensitivity of human T lymphocytes assessed by the colony-forming assay. Radiat Res 120: 164–176
Gunning P, Leavitt J, Muscat G, Ng SY and Kedes L (1987) A human beta-actin expression vector system directs high-level accumulation of antisense transcripts. Proc Natl Acad Sci USA 84: 4831–4835
Hietanen T, Kellokumpu-Lehtinen P and Pitkanen M (1995) Action of recombinant interferons and interleukin 2 in modulating radiation effects on viability and cytotoxicity of large granular lymphocytes. Int J Radiat Biol 67: 119–126
Hill RP (1986) Sensitizers and radiation dose fractionation: results and interpretations. Int J Radiat Oncol Biol Physics 12: 1049–1054
Hockel M, Knoop C, Schlenger K, Vorndran B, Baussmann E, Mitze M, Knapstein PG and Vaupel P (1993) Intratumoral pO2predicts survival in advanced cancer of the uterine cervix. Radiother Oncol 26: 45–50
Howell RL and Koch CJ (1980) The disaggregation, separation and identification of cells from irradiated and unirradiated EMT6 mouse tumors. Int J Radiat Oncol Biol Physics 6: 311–318
Jaffee EM and Pardoll DM (1997) Considerations for the clinical development of cytokine gene-transduced tumor cell vaccines. Methods 12: 143–153
James SE, Arlett CF, Green MH and Bridges BA (1983) Radiosensitivity of human T-lymphocytes proliferating in long term culture. Int J Radiat Biol Related Studies Physics, Chem Med 44: 417–422
Koch CJ, Evans SM and Lord EM (1995) Oxygen dependence of cellular uptake of EF5 [2-(2-nitro-1H-imidazol-1-yl)-N-(2,2,3,3,3-pentafluoropropyl)acetamide]: analysis of drug adducts by fluroescent antibodies vs bound radioactivity. Br J Cancer 72: 869–874
Kwok TT and Sutherland RM (1992) Cell cycle dependence of epidermal growth factor induced radiosensitization. Int J Radiat Oncol Biol Physics 22: 525–527
Ledbetter JA and Herzenberg LA (1979) Xenogeneic monoclonal antibodies to mouse lymphoid differentiation antigens. Immunol Rev 47: 63–90
Lee J, Fenton BM, Koch CJ, Frelinger JG and Lord EM (1998) Interleukin 2 expression by tumor cells alters both the immune response and the tumor microenvironment. Cancer Res 58: 1478–1485
Lee J, Siemann DW, Koch CJ and Lord EM (1996) Direct relationship between radiobiological hypoxia in tumors and monoclonal antibody detection of EF5 cellular adducts. Int J Cancer 67: 372–378
McAdam AJ, Felcher A, Woods ML, Pulaski BA, Hutter EK, Frelinger JG and Lord EM (1994) Transfection of transforming growth factor-beta producing tumor EMT6 with interleukin-2 elicits tumor rejection and tumor reactive cytotoxic T-lymphocytes. J Immunother Emphasis Tumor Immunol 15: 155–164
McAdam AJ, Pulaski BA, Storozynsky E, Yeh KY, Sickel JZ, Frelinger JG and Lord EM (1995) Analysis of the effect of cytokines (interleukins 2, 3, 4, and 6, granulocyte-monocyte colony-stimulating factor, and interferon-gamma) on generation of primary cytotoxic T lymphocytes against a weakly immunogenic tumor. Cell Immunol 165: 183–192
McBride WH, Economou JS, Kuber N, Hong JH, Chiang CS, Syljuasen R, Dougherty ST and Dougherty GJ (1995) Modification of tumor microenvironment by cytokine gene transfer. Acta Oncol 34: 447–451
Miller AR, McBride WH, Hunt K and Economou JS (1994) Cytokine-mediated gene therapy for cancer. Ann Surg Oncol 1: 436–450
Mor F and Cohen IR (1996) IL-2 rescues antigen-specific T cells from radiation or dexamethasone-induced apoptosis. Correlation with induction of Bcl-2. J Immunol 156: 515–522
Moulder JE and Rockwell S (1984) Hypoxic fractions of solid tumors: experimental techniques, methods of analysis, and a survey of existing data. Int J Radiat Oncol Biol Physics 10: 695–712
Pulaski BA, Yeh KY, Shastri N, Maltby KM, Penney DP, Lord EM and Frelinger JG (1996) Interleukin 3 enhances cytotoxic T lymphocyte development and class I major histocompatibility complex ‘re-presentation’ of exogenous antigen by tumor-infiltrating antigen-presenting cells. Proc Natl Acad Sci USA 93: 3669–3674
Seki H, Iwai K, Kanegane H, Konno A, Ohta K, Ohta K, Yachie A, Taniguchi N and Miyawaki T (1995) Differential protective action of cytokines on radiation-induced apoptosis of peripheral lymphocyte subpopulations. Cell Immunol 163: 30–36
Syljuasen RG, Belldegrun A, Tso CL, Withers HR and McBride WH (1997) Sensitization of renal carcinoma to radiation using alpha interferon (IFNA) gene transfection. Radiat Res 148: 443–448
Yokota T, Arai N, Lee F, Rennick D, Mosmann T and Arai K (1985) Use of a cDNA expression vector for isolation of mouse interleukin 2 cDNA clones: expression of T-cell growth-factor activity after transfection of monkey cells. Proc Natl Acad Sci USA 82: 68–72
Author information
Authors and Affiliations
Rights and permissions
From twelve months after its original publication, this work is licensed under the Creative Commons Attribution-NonCommercial-Share Alike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
About this article
Cite this article
Lee, J., Moran, J., Fenton, B. et al. Alteration of tumour response to radiation by interleukin-2 gene transfer. Br J Cancer 82, 937–944 (2000). https://doi.org/10.1054/bjoc.1999.1022
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1054/bjoc.1999.1022
Keywords
This article is cited by
-
Recent developments in oncolytic adenovirus-based immunotherapeutic agents for use against metastatic cancers
Cancer Gene Therapy (2013)
-
Adequate Antigen Availability: A Key Issue for Novel Approaches to Tumor Vaccination and Tumor Immunotherapy
Journal of Neuroimmune Pharmacology (2013)
-
Optimal MHC-II-restricted tumor antigen presentation to CD4+ T helper cells: the key issue for development of anti-tumor vaccines
Journal of Translational Medicine (2012)
-
The Tumor-Immune Microenvironment and Response to Radiation Therapy
Journal of Mammary Gland Biology and Neoplasia (2010)
-
The hepatic transcriptome in human liver disease
Comparative Hepatology (2006)