Abstract
RENCA-IL-2 (Murine Renal Cell Carcinoma transfected with murine IL-2 gene) cells were rejected by immunocompetent (but not T-cell deficient) Balb/c mice, which developed ‘immunity’ to subsequent parental RENCA tumour cell challenge. Splenocytes adoptively transferred this immunity. CD4+ and CD8+ T-lymphocytes prepared from the spleens of ‘tumour immune’ mice were evaluated for their ability to traffic into the tumour environment using an in vivo model that enables visualization of events within the microvasculature. RENCA cells were implanted into the mouse cremaster muscle and the trafficking of syngeneic lymphocyte subpopulations, derived from naive and ‘immune’ animals, into both the RENCA tumour and the surrounding normal cremaster muscle microcirculation was measured by in vivo microscopy. Fluorescently labelled CD4+ and CD8+ T lymphocytes taken from the spleens of naive mice or mice previously immunized with RENCA-IL-2 were injected systemically into tumour-bearer mice. Naive effector cells migrated to, and flowed through both the tumour and the normal microcirculation, with negligible adhesion. However we observed the selective recruitment, localization and arrest of immune CD4+ and CD8+ T lymphocytes (P < 0.05) into the tumour microcirculation, and in some instances the subsequent extravasation of cells into the tumour interstitium. Lymphocyte rolling by ‘immune’ CD4+ and CD8+ T-cells in the tumour microcirculation was greatly reduced, suggesting impaired adhesion molecule expression on the tumour endothelium. This study clearly demonstrates, by direct in vivo microscopy assessment, the localization of effector cells, CD4+ and CD8+ lymphocytes into 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
Adams DH, Yannelli JR, Mnewman W, Lawley T, Ades E, Rosenber SA and Shaw S (1997) Adhesion of tumour-infiltrating lymphocytes to endothelium: a phenotypic and functional analysis. Br J Cancer 75: 1421–1431
Ali SA, Mclean CS, Boursnell MEG, Martin G, Holmes L, Reeder S, Entwisle C, Blakeley DM, Todryk S, Vile R, Robins RA and Rees RC (2000) The pre-clinical evaluation of ‘whole’ cell vaccines for prophylaxis and therapy: the use of DISC-HSV vector to transduce cytokine genes. Cancer Res 60: 1663–1670
Basse PH, Nanmark U, Johansson BR, Herberman RB and Goldfarb RH (1991) Establishment of cell-to-cell contacts by adoptively transferred adherent lymphokine activated killer cells with metastatic melanoma cells. J Natl Cancer Inst 83: 944–950
Bassil B, Dosoretz DE and Prout, Jr GR (1985) Validation of the tumour, nodes and metastasis classification of renal cell carcinoma. J Urol 134: 450–454
Bear HD (1986) Tumour specific suppressor T cells which inhibit the in vitro generation of cytotoxic T cells from immune and early tumour-bearing host spleens. Cancer Res 46: 1805–1812
Blay JY, Negrier S, Combaret V, Attali S, Goillot E, Merrouche Y, Mercatello A, Ravault A, Tourani JM, Moskovtchenko JF, Philip T and Favrot M (1992) Serum levels of IL-6 as a prognostic factor in metastatic renal cell carcinoma. Cancer Res 52: 3317–3322
Bradley LM and Watson SR (1996) Lymphocyte migration into tissue; the paradigm derived from CD4 subsets. Current Opinions in Immunology 8: 312
Brown NJ and Reed MWR (1997) Leucocyte interactions with the mouse cremaster muscle microcirculation in vivo in response to tumour conditioned medium. Br J Cancer 75: 993–999
Brown NJ, Ali S, Reed MWR, Wiltrout R and Rees RC (1997) Trafficking of activated lymphocytes into the RENCA tumour microcirculation in vivo in mice. Br J Cancer 76: 1572–1578
Cavallo F, Giovarelli M, Gulino A, Vacca A, Stoppacciaro A, Modesti A and Forni G (1992) Role of neutrophils and CD4+ lymphocytes in the primary and memory response to non-immunogenic murine mammary adenocarcinoma made immunogenic by IL-2 gene. J Immunol 149: 3627–3635
Colombo MP and Forni G (1996) Immunotherapy 1: cytokine gene transfer strategies. Cancer Metastasis Reviews 15: 317–328
Fakhrai H, Shawler DL and Gjerset Ret al (1995) Cytokine gene therapy with interleukin 2 transduced fibroblasts: effect of IL-2 dose on anti-tumour immunity. Human Gene Therapy 6: 591–601
Figlin R, Gitlitz B, Franklin J, Dorey F, Moldawer N, Rausch J, Dekernion J and Belldegrun A (1997) Interleukin-2 based immunotherapy for the treatment of metastatic renal cell carcinoma: an anlysis of 203 consecutively treated patients. Cancer J Sci Am 3: S92–97
Fukumura D, Salehi HA, Witwer B, Tuma RF, Melder RI and Jain Rk-K (1995) Tumour necrosis factor induced leucocyte adhesion in normal and tumour vessels: effect of tumour type, transplantation site and host strain. Cancer Res 55: 4824–4829
Fumagalli L, Lissoni P, Di Felice G, Meregalli S, Valsuani G, Mengo S and Rovelli F (1999) Pretreatment serum markers and lymphocyte response to interleukin therapy. Br J Cancer 80: 407–411
Garbett EA, Reed MWR and Brown NJ (1994) Viability and visualisation of fluorescently labelled lymphocytes in vitro and in vivo. Int J Micro Clin Exp 14: 249
Jicha DL, Mule JJ and Rosenberg SA (1991) Interleukin 7 generates antitumour cytotoxic T lymphocytes against murine sarcomas with efficacy in cellular adoptive immunotherapy. J Exp Med 174: 1511–1515
Kahn M, Sugawara H, Macgowan P, Okuno K, Nagoya S, Hellstorm KE, Hellstrom I and Greenberg P (1991) CD4+ T cell clones specific for human p97 melanoma-associated antigen can eradicate pulmonary metastases from a murine tumour expressing the p97 antigen. J Immunol 146: 3235–3241
Karasuyama H and Melchers F (1988) Establishmentof mouse cell lines which constitutively secrete large quantities of interleukin 2, 3, 4, or 5, using modified cDNA expression vectors. Eur J Immunol 18: 97–104
Kern DE, Klarnet JP, Jensen MC and Greenberg PD (1986) Requirement for recognition of class II molecules and processed tumour antigen for optimal generation of syngeneic tumour-specific class I restricted CTL. J Immunol 136: 4303–4310
Kradin RL, Kurnick JT and Lazarus DSet al (1989) Tumour infiltrating lymphocytes and interleukin-2 in the treatment of advanced cancer. The Lancet 1: 577–580
Ley K, Bullard DC, Arbones ML, Bosse R, Vestweber D, Tedder TF and Beaudet AI (1995) Sequential contribution of L- and P-selectin to leucocyte rolling in vivo. J Exp. Med 181: 669–675
Litton MI, Dohlsten M, Rosendahl A, Ohlsson L, Sogaard M, Anderssoon J and Andersson U (1999) The distinct role of CD4+ and CD8+ T-cells during the anti-tumour effects of targeted superantigens. Brit J Cancer 81: 359–366
Lopez-Hanninen E, Kirchner H and Atzpodien J (1996) IL-2 based home therapy of metastatic renal cell carcinoma: risks and benefits in 215 consecutive single institution patients. J Urol 155: 19–25
Lynch DH and Miller RE (1991) Immunotherapeutic elimination of syngeneic tumours in vivo by cytotoxic T lymphocytes generated in vitro from lymphocytes from draining lymph nodes of tumour bearing mice. Eur J Immunol 21: 1403–1410
McAdam AJ, Pulsaki BA and Storozynsky Eet al (1995) Analysis of effect of cytokines (interleukins 2, 3, 4, and 6, Granulocyte-Monocyte Colony Stimulating Factor, and interferon-γ) on generation of primary cytotoxic T lymphocytes against a weakly immunogenic tumour. Cellular Immunol 165: 183–192
McKinnon JG, Lalani A and Bear HD (1993) Generation of cytotoxic T lymphocytes from peripheral blood. Surgery 113: 536–540
Melder RI, Salehi HA and Jain RK (1995) Interaction of activated natural killer cells with normal and tumour vessels in cranial windows in mice. Microvasc Res 50: 35–44
Murphy GP and Hruskesky WI (1980) A murine renal cell carcinoma. J Natl Cancer Inst 50: 1013
Palmer PA, Vinke J, Evers P, Pourreau C, Oskram R, Roest G, Vlems F, Becker L, Loriaux E and Franks CR (1992) Continuous infusion of recombinant interleukin-2 with or without autologous lymphokine activated killer cells for the treatment of advanced renal cell carcinoma. Europ J Cancer 28A: 1038–1044
Pawelec G, Rees RC, Kiessling R, Madrigal A, Dodi A, Baxevanis C, GambacortiPasserini C, Masucci G and Zeuthen J (1999) Cells and cytokines in immunotherapy and gene therapy of cancer. Critical Reviews in Oncogenesis 10: 83–127
Rees RC and Mian S (1999) Selective MHC expression in tumours modulates adaptive and innate antitumour responses. Cancer Immunol Immunother 48: 374–381
Rosenberg SA (1991) Immunotherapy and gene therapy of cancer. Cancer Res 51, (Suppl)5074–5079
Rosenberg SA (1995) Treatment of patients with metastatic melanoma with autologous tumor-infiltrating lymphocytes and interleukin-2. J Natl Cancer Inst 87: 319
Rosenberg SA, Packard BS and Aebersold PM (1988) Use of tumour infiltrating lymphocytes and IL2 in immunotherapy of patients with metastatic melanoma. New Eng J Med 319: 1676–1680
Sasaki A, Melder RI, Whiteside TL, Herberman RB and Jain RK (1991) Preferential localisation of human adherent lymphokine-activated killer cells in tumour microcirculation. J Natl Cancer Inst 83: 433–437
Sayers TI, Brooks AD, Lee JK, Fenton RG, Komschlies KI, Wigginton JM, Winkler-Pickett R and Wiltrout RH (1998) Molecular mechanisms of immune-mediated lysis of murine renal cancer: differential contributions of perforin-dependent versus Fas-mediated pathways in lysis by NK and T cells. J Immunol 161: 3957–3965
Schoof DD, Douville L, Terashima Y, Richie JP and Batter S Eberleintj (1993) Survival characteristics of metastatic renal cell carcinoma patients treated with lymphokine-activated killer cells plus interleukin-2. Urology 41: 534–539
Sedlmayr P, Rabinowich H, Elder E, Ernstoff MS, Kirkwood JM, Herberman RB and Whiteside T (1991) Depressed ability of patients with melanoma or renal cell carcinoma to generate adherent lymphokine activated killer cells. J Immunotherapy 10: 336–346
Springer TA (1994) Traffic signals for lymphocyte recirculation and leucocyte emigration; the multistep paradigm. Cell 76: 301
Tepper RI and Mule JI (1994) Experimental and clinical studies of cytokine-gene modified tumour cell. Human Gene Therapy 5: 153–164
Zaks TZ, Chappell DB, Rosenberg SA and Restifo NP (1999) Fas-mediated suicide of tumour-reactive T cells following activation by specific tumour: selective rescue by caspase inhibition. J Immunol 162: 3273–3279
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
Ali, S., Rees, R., Anderson, D. et al. Trafficking of ‘immune’ CD4+/CD8+ T-lymphocytes into the RENCA tumour microcirculation in vivo in mice. Br J Cancer 83, 1061–1068 (2000). https://doi.org/10.1054/bjoc.2000.1403
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1054/bjoc.2000.1403
Keywords
This article is cited by
-
Intravital imaging of anti-tumor immune response and the tumor microenvironment
Seminars in Immunopathology (2010)
-
Immunotherapeutic potential of DISC-HSV and OX40L in cancer
Cancer Immunology, Immunotherapy (2006)