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Annals of Surgical Oncology

, Volume 3, Issue 2, pp 176–184 | Cite as

In situ cytokine production by breast cancer tumor-infiltrating lymphocytes

  • Benjamin J. Camp
  • Sonya T. Dyhrman
  • Vincent A. Memoli
  • Leila A. Mott
  • Richard J. BarthJr
Original Articles

Abstract

Background: Human breast cancers progressively grow despite the presence of extensive lymphocytic infiltration and specific antitumor immune recognition, thereby calling into question the competency of breast tumor-infiltrating lymphocytes (TIL). The function of breast TILs in vivo and their possible role in the suppression of an antitumor immune response are largely unknown.

Methods: The cytokines produced in situ by lymphocytes in 89 breast carcinomas and 14 benign breast lesions were assessed using immunohistochemistry.

Results: The majority of tumor and benign breast samples contained T-cell infiltrates, which were disclosed using an anti-CD3 antibody stain. The percentage of tumor samples in which ⩾3% of the lymphocytes were producing cytokines was as follows: interleukin (IL)-2 45%, IL-4 36%, tumor necrosis factor-alpha (TNF-α) 28%, transforming growth factor-beta 1 (TGF-β1) 20%, IL-10 11%, interferon-gamma (IFN-γ) 4%, and granulocytemacrophage colony-stimulating factor (GM-CSF) 3%. Production of IL-2, IL-4, and TGF-β1 by TILs in breast cancers exceeded that detected in benign breast lesions (p<0.005). Significantly more tumor samples contained lymphocytes producing IL-2, IL-4, TGF-β1, and TNF-α than IFN-γ and GM-CSF (p<0.002 for each comparison). One or more of the potentially immunoinhibitory cytokines—IL-4, IL-10, or TGF-β1—were produced by lymphocytes in 44% of the specimens. No significant associations were seen between lymphocyte production of a particular cytokine and disease-free survival (median follow-up 43 months).

Conclusions: Immunohistochemical techniques can be used to detect cytokine secretion by TILs in preserved tissue. The relative lack of secretion of IFN-γ and GM-CSF, rather than a deficiency of IL-2, may explain why the antitumor immune response to breast cancer is impaired.

Key Words

Breast cancer Tumor-infiltrating lymphocyte Immunohistochemistry Interleukin-2 Interleukin-4 Tumor growth factor-beta 1 

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References

  1. 1.
    Eremin O, Coombs RA, Prospero TA, Plumb D. T-lymphocyte subpopulations infiltrating human mammary carcinomas.J Natl Cancer Inst 1982;69:1–8.PubMedGoogle Scholar
  2. 2.
    Shimokawara I, Imamura M, Yamanaka N, Ishii Y, Kikuchi K. Identification of lymphocyte subpopulations in human breast cancer tissue and its significance: an immunoperoxidase study with anti-human T- and B-cell sera.Cancer 1982;49:1456–64.PubMedGoogle Scholar
  3. 3.
    Wintzer H, Bohle W, von Kleist S. Study of the relationship between immunohistologically demonstrated lymphocytes infiltrating human breast carcinomas and patients' survival.Can Res Clin Oncol 1991;117:163–7.Google Scholar
  4. 4.
    Whitford P, Mallon EA, George WD, Campbell AM. Flow cytometric analysis of tumor infiltrating lymphocytes in breast cancer.Br J Can 1990;62:971–5.Google Scholar
  5. 5.
    Schwartzentruber DJ, Solomon D, Rosenberg SA, Topalian SL. Characterization of lymphocytes infiltrating human breast cancer: specific immune reactivity detected by measuring cytokine secretion.J Immunother 1992;12:1–12.PubMedGoogle Scholar
  6. 6.
    Miescher S, Whiteside TL, Carrel S, von Fliedner V. Functional properties of tumor-infiltrating and blood lymphocytes in patients with solid tumors: effects of tumor cells and their supernatants on proliferative responses of lymphocytes.J Immunol 1986;136:1899–907.PubMedGoogle Scholar
  7. 7.
    Miescher S, Stoeck M, Qiao L, Barras C, Barrelet L, von Fliedner V. Preferential clonogenic deficit of CD8-positive T-lymphocytes infiltrating human solid tumors.Can Res 1988;48:6992–8.Google Scholar
  8. 8.
    Vose BM, Moore M. Supressor cell activity of lymphocytes infiltrating human lung and breast tumours.Int J Cancer 1979;24:579–85.PubMedGoogle Scholar
  9. 9.
    Asher AL, Mulé JJ, Kasid A, et al. Murine tumor cells transduced with the gene for tumor necrosis factor-α: evidence for paracrine immune effects of tumor necrosis factor against tumors.J Immunol 1991;146:3227–34.PubMedGoogle Scholar
  10. 10.
    Gansbacher B, Bannerji R, Daniels B, Zier K, Cronin K, Gilboa E. Retroviral vector-mediated γ interferon gene transfer into tumor cells generates potent and long lasting antitumor immunity.Can Res 1990;50:7820–5.Google Scholar
  11. 11.
    Fearon ER, Pardoll DM, Itaya T, et al. Interleukin 2 production by tumor cells bypasses T helper function in the generation of an antitumor response.Cell 1990;60:397–403.CrossRefPubMedGoogle Scholar
  12. 12.
    Dranoff G, Jaffel E, Lazenby A, et al. Vaccination with irradiated tumor cells engineered to secrete murine GM-CSF stimulates potent, specific, and long lasting anti-tumor immunity.Proc Natl Acad Sci U S A 1993;90:3539–43.PubMedGoogle Scholar
  13. 13.
    Barth RJ, Mulé JJ, Spiess P, Rosenberg SA. Interferon γ and tumor necrosis factor have a role in tumor regressions mediated by murine tumor infiltrating lymphocytes.J Exp Med 1991;173:647–58.CrossRefPubMedGoogle Scholar
  14. 14.
    Pisa P, Halapi E, Pisa EK, et al. Selective expression of interleukin 10, interferon γ, and granulocyte-macrophage colony-stimulating factor in ovarian cancer biopsies.Proc Natl Acad Sci U S A 1992;89:7708–12.PubMedGoogle Scholar
  15. 15.
    deWaal Malefyt R, Yssel H, deVries JE. Direct effects of IL-10 on subsets of human CD4+ T cell clones and resting T cells.J Immunol 1993;150:4754–65.Google Scholar
  16. 16.
    Wang P, Wu P, Siegel MI, Egan RW, Billah MM. IL-10 inhibits transcription of cytokine genes in human peripheral blood mononuclear cells.J Immunol 1994;153:811–6.PubMedGoogle Scholar
  17. 17.
    Schlaak JF, Hermann E, Gallati H, Zum Buschenfelde KH, Fleischer B. Differential effects of IL-10 on proliferation and cytokine production of human γ/δ and α/β T cells.Scand J Immunol 1994;39:209–15.PubMedGoogle Scholar
  18. 18.
    deWaal Malefyt R, Abrams JS, Bennet BJ, Figdor CG, de Vries JE. IL-10 inhibits cytokine synthesis by human monocytes: an autoregulatory role of IL-10 produced by monocytes.J Exp Med 1991;174:1209–20.Google Scholar
  19. 19.
    Salgame P, Abrams JS, Clayberger C, et al. Differing lymphokine profile of functional subsets of human CD4 and CD8 T cell clones.Science 1991;254:279–82.PubMedGoogle Scholar
  20. 20.
    Martinez O, Gibbons R, Garavoy FJ, Aronson FR. IL-4 inhibits IL-2 receptor expression and IL-2 dependent proliferation on human T cells.J Immunol 1990;144:2211–5.PubMedGoogle Scholar
  21. 21.
    Chen Y, Kuchroo V, Inobe J, Hafler D, Weiner H. Regulatory T cell clones induced by oral tolerance: suppression of autoimmune encephalomyelitis.Science 1994;265:1237–40.PubMedGoogle Scholar
  22. 22.
    Kehrl JH, Wakefield LM, Roberts AB, et al. Production of transforming growth factor B by human T lymphocytes and its potential role in the regulation of T cell growth.J Exp Med 1986;163:1037–50.CrossRefPubMedGoogle Scholar
  23. 23.
    Elston CW, Gresham GA, Rao GS, et al. The cancer research campaign (King's/Cambridge) trial for early breast cancer: clinico-pathological aspects.Br J Cancer 1982;45:655–69.PubMedGoogle Scholar
  24. 24.
    Dawson PJ, Karrsion T, Ferguson DJ. Histologic features associated with long term survival in breast cancer.Hum Pathol 1986;17:1015–21.PubMedGoogle Scholar
  25. 25.
    Rosen PP, Saigo PE, Braun DW, Weathers E, DePalo A. Predictors of recurrence in stage I (T1NOMO) breast carcinoma.Ann Surg 1981;193:15–25.PubMedGoogle Scholar
  26. 26.
    Tanaka M, Tanaka H, Ishikawa E. Immunohistochemical demonstration of surface antigen of human lymphocytes with monoclonal antibody in acetone-fixed paraffin-embedded sections.J Histochem Cytochem 1984;32:452–4.PubMedGoogle Scholar
  27. 27.
    Sato Y, Mukai K, Watanabe S, Goto M, Shimosato Y. The AMeX method: a simplified technique of tissue processing and paraffin embedding with improved preservation of antigens for immunostaining.Am J Pathol 1986;125:431–5.PubMedGoogle Scholar
  28. 28.
    Hoefakker S, van t Erve EHM, Deen C, et al. Immunohistochemical detection of co-localizing cytokine and antibody producing cells in the extrafollicular area of human palatine tonsils.Clin Exp Immunol 1993;93:223–8.PubMedGoogle Scholar
  29. 29.
    Pickvance EA, Oegema TR, Thompson RC. Immunolocalization of selected cytokines and proteases in canine articular cartilage after transarticular loading.J Orthop Res 1992;11:313–23.Google Scholar
  30. 30.
    Vitolo D, Zerbe T, Kanbour A, Dahl C, Herberman RB, Whiteside TL. Expression of mRNA for cytokines in tumor-infiltrating mononuclear cells in ovarian adenocarcinoma and invasive breast cancer.Int J Cancer 1992;51:573–80.PubMedGoogle Scholar
  31. 31.
    Schoof DD, Terashima Y, Peoples GP, et al. CD4+ T cell clones isolated from human renal cell carcinoma possess the functional characteristics of Th2 helper cells.Cell Immun 1993;150:114–23.CrossRefGoogle Scholar
  32. 32.
    Whiteside TL, Miescher S, Hurlimann J, Moretta L, von Fliedner. Clonal analysis and in situ characterization of lymphocytes infiltrating human breast carcinomas.Can Immun Immunother 1986;23:169–78.Google Scholar
  33. 33.
    Alexander JP, Kudoh S, Melsop KA, et al. T cells infiltrating renal cell carcinoma display a poor proliferative response even though they can produce interleukin 2 and express interleukin 2 receptors.Can Res 1993;53:1380–7.Google Scholar
  34. 34.
    Finke J, Zea A, Stanley J, et al. Loss of T cell receptor z chain and p56lck in T cells infiltrating human renal cell carcinoma.Can Res 1993;53:5613–6.Google Scholar
  35. 35.
    Lombardi G, Sidhu S, Batchelor R, Lechler R. Anergic T cells as suppressor cells in vitro.Science 1994;264:1587–9.PubMedGoogle Scholar
  36. 36.
    McCume BK, Mullin BR, Flanders KC, Jaffurs WJ, Mullen LT, Sporn MB. Localization of transforming growth factor-beta isotypes in lesions of the human breast.Hum Pathol 1992;23:13–20.Google Scholar
  37. 37.
    Gorsch SM, Memoli VA, Stukel TA, Gold LI, Arrick BA. Immunohistochemical staining for transforming growth factor b1 associates with disease progression in human breast cancer.Can Res 1992;52:6949–52.Google Scholar
  38. 38.
    Walker RA, Dearing SJ, Gallacher B. Relationship of transforming growth factor beta 1 to extracellular matrix and stromal infiltrates in invasive breast carcinoma.Br J Cancer 1994;69:1160–5.PubMedGoogle Scholar
  39. 39.
    Murray PA, Barrett-Lee P, Travers M, Luqmani Y, Powles T, Coombes RC. The prognostic significance of transforming growth factors in human breast cancer.Br J Cancer 1993;67:1408–12.PubMedGoogle Scholar
  40. 40.
    Hwu P, Yannelli J, Kriegler M, et al. Functional and molecular characterization of tumor-infiltrating lymphocytes transduced with TNF-α cDNA for the gene therapy of cancer in humans.J Immunol 1993;150:4104–15.PubMedGoogle Scholar

Copyright information

© The Society of Surgical Oncology, Inc. 1996

Authors and Affiliations

  • Benjamin J. Camp
    • 1
  • Sonya T. Dyhrman
    • 4
  • Vincent A. Memoli
    • 2
  • Leila A. Mott
    • 3
  • Richard J. BarthJr
    • 1
  1. 1.From the Department of SurgeryDartmouth-Hitchcock Medical Center and Norris Cotton Cancer CenterLebanonUSA
  2. 2.the Department of PathologyDartmouth-Hitchcock Medical Center and Norris Cotton Cancer CenterLebanonUSA
  3. 3.the Department of Community and Family MedicineDartmouth-Hitchcock Medical Center and Norris Cotton Cancer CenterLebanonUSA
  4. 4.the Department of BiologyDartmouth-Hitchcock Medical Center and Norris Cotton Cancer CenterLebanonUSA

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