Immune dysfunction and micrometastases in women with breast cancer

  • Michael J. Campbell
  • Janet Scott
  • Holden T. Maecker
  • John W. Park
  • Laura J. Esserman


Cytokines produced by T lymphocytes are critical to the efficacy of a given immune response and dysregulation of immune responses may play a role in cancer progression. We assessed the intracellular cytokine profiles of T cells in the peripheral blood of women with breast cancer and explored the relationship of these responses with the presence of cancer in lymph nodes and bone marrow. Peripheral blood lymphocytes from 84 patients and 26 healthy volunteers were analyzed by 4-color flow cytometry for surface markers and for intracellular cytokines. Bone marrow samples from some of these patients were also collected and analyzed for the presence of epithelial cells (micrometastases) by flow cytometry. The percentages of both CD4+ and CD8+ cells producing type1 (IL-2, IFN-γ or TNF-α) and type 2 (IL-4) were significantly lower in patients with breast cancer compared to healthy controls. These results indicate a general immune dysfunction in these patients as opposed to a shift in the balance of type1 and type2 cells. These dysregulated T cell responses did not correlate with age, stage of disease, or nodal status. However, we did observe a correlation between number of micrometastases in the bone marrow and T cell responsiveness.


breast cancer cytokines flow cytometry immune dysfunction 


  1. 1.
    Pellegrini, P, Berghella, AM, Del Beato, T, Cicia, S, Adorno, D, Casciani, CU 1996Disregulation in TH1 and TH2 subsets of CD4+ T cells in peripheral blood of colorectal cancer patients and involvement in cancer establishment and progressionCancer Immunol Immunother4218CrossRefPubMedGoogle Scholar
  2. 2.
    Angevin, E, Kremer, F, Gaudin, C, Hercend, T, Triebel, F 1997Analysis of T-cell immune response in renal cell carcinoma: polarization to type 1-like differentiation pattern, clonal T-cell expansion and tumor-specific cytotoxicityInt J Cancer72431440CrossRefPubMedGoogle Scholar
  3. 3.
    Sato, M, Goto, S, Kaneko, R, Ito, M, Sato, S, Takeuchi, S 1998Impaired production of Th1 cytokines and increased frequency of Th2 subsets in PBMC from advanced cancer patientsAnticancer Res1839513955PubMedGoogle Scholar
  4. 4.
    Goto, S, Sato, M, Kaneko, R, Itoh, M, Sato, S, Takeuchi, S 1999Analysis of Th1 and Th2 cytokine production by peripheral blood mononuclear cells as a parameter of immunological dysfunction in advanced cancer patientsCancer Immunol Immunother48435442CrossRefPubMedGoogle Scholar
  5. 5.
    Tabata, T, Hazama, S, Yoshino, S, Oka, M 1999Th2 subset dominance among peripheral blood T lymphocytes in patients with digestive cancersAm J Surg177203208CrossRefPubMedGoogle Scholar
  6. 6.
    Ito, N, Nakamura, H, Tanaka, Y, Ohgi, S 1999Lung carcinoma: analysis of T helper type 1 and 2 cells and T cytotoxic type 1 and 2 cells by intracellular cytokine detection with flow cytometryCancer8523592367CrossRefPubMedGoogle Scholar
  7. 7.
    Zhang, XL, Komada, Y, Chipeta, J, Li, QS, Inaba, H, Azuma, E, Yamamoto, H, Sakurai, M 2000Intracellular cytokine profile of T cells from children with acute lymphoblastic leukemiaCancer Immunol Immunother49165172CrossRefPubMedGoogle Scholar
  8. 8.
    Mosmann, TR, Coffman, RL 1989TH1 and TH2 cells: different patterns of lymphokine secretion lead to different functional propertiesAnnu Rev Immunol7145173CrossRefPubMedGoogle Scholar
  9. 9.
    Romagnani, S 1991Human TH1 and TH2 subsets: doubt no moreImmunol Today12256257CrossRefPubMedGoogle Scholar
  10. 10.
    Lucey, DR, Clerici, M, Shearer, GM 1996Type 1 and type 2 cytokine dysregulation in human infectious, neoplastic, and inflammatory diseasesClin Microbiol Rev9532562PubMedGoogle Scholar
  11. 11.
    Romagnani, S 1997The Th1/Th2 paradigmImmunol Today18263266CrossRefPubMedGoogle Scholar
  12. 12.
    Romagnani, S, Parronchi, P, D’Elios, MM, Romagnani, P, Annunziato, F, Piccinni, MP, Manetti, R, Sampognaro, S, Mavilia, C, De Carli, M, Maggi, E, Del Prete, GF 1997An update on human Th1 and Th2 cellsInt Arch Allergy Immunol113153156PubMedGoogle Scholar
  13. 13.
    Punnonen, R, Teisala, K, Kuoppala, T, Bennett, B, Punnonen, J 1998Cytokine production profiles in the peritoneal fluids of patients with malignant or benign gynecologic tumorsCancer83788796PubMedGoogle Scholar
  14. 14.
    Onishi, T, Ohishi, Y, Imagawa, K, Ohmoto, Y, Murata, K 1999An assessment of the immunological environment based on intratumoral cytokine production in renal cell carcinomaBJU Int83488492PubMedGoogle Scholar
  15. 15.
    Lauerova, L, Dusek, L, Simickova, M, Kocak, I, Vagundova, M, Zaloudik, J, Kovarik, J 2002Malignant melanoma associates with Th1/Th2 imbalance that coincides with disease progression and immunotherapy responseNeoplasma49159166PubMedGoogle Scholar
  16. 16.
    Jung, T, Schauer, U, Heusser, C, Neumann, C, Rieger, C 1993Detection of intracellular cytokines by flow cytometryJ Immunol Methods159197207PubMedGoogle Scholar
  17. 17.
    Picker, LJ, Singh, MK, Zdraveski, Z, Treer, JR, Waldrop, SL, Bergstresser, PR, Maino, VC 1995Direct demonstration of cytokine synthesis heterogeneity among human memory/effector T cells by flow cytometryBlood8614081419PubMedGoogle Scholar
  18. 18.
    Maino, VC, Picker, LJ 1998Identification of functional subsets by flow cytometry: intracellular detection of cytokine expressionCytometry34207215PubMedGoogle Scholar
  19. 19.
    Suni, MA, Picker, LJ, Maino, VC 1998Detection of antigen-specific T cell cytokine expression in whole blood by flow cytometryJ Immunol Methods2128998PubMedGoogle Scholar
  20. 20.
    Maecker, HT, Maino, VC, Picker, LJ 2000Immunofluorescence analysis of T-cell responses in health and diseaseJ Clin Immunol20391399PubMedGoogle Scholar
  21. 21.
    Ghanekar, SA, Maecker, HT 2003Cytokine flow cytometry: multiparametric approach to immune function analysisCytotherapy516PubMedGoogle Scholar
  22. 22.
    Akimoto, M, Ishii, H, Nakajima, Y, Iwasaki, H, Tan, M, Abe, R, Kasai, M 1986Assessment of host immune response in breast cancer patientsCancer Detect Prev9311317PubMedGoogle Scholar
  23. 23.
    Head, JF, Elliott, RL, McCoy, JL 1993Evaluation of lymphocyte immunity in breast cancer patientsBreast Cancer Res Treat267788PubMedGoogle Scholar
  24. 24.
    Wiltschke, C, Krainer, M, Budinsky, AC, Berger, A, Muller, C, Zeillinger, R, Speiser, P, Kubista, E, Eibl, M, Zielinski, CC 1995Reduced mitogenic stimulation of peripheral blood mononuclear cells as a prognostic parameter for the course of breast cancer: a prospective longitudinal studyBr J Cancer7112921296PubMedGoogle Scholar
  25. 25.
    Nicolini, A, Ferrari, P, Spinelli, R, Carpi, A, Sagripanti, A, Ambrogi, F 1996Cell-mediated immunity in breast cancer patientsBiomed Pharmacother50337343PubMedGoogle Scholar
  26. 26.
    Mccoy, JL, Rucker, R, Petros, JA 2000Cell-mediated immunity to tumor-associated antigens is a better predictor of survival in early stage breast cancer than stage, grade or lymph node statusBreast Cancer Res Treat60227234PubMedGoogle Scholar
  27. 27.
    Nicolini, A, Carpi, A, Ferrari, P, Tartarelli, G, Anselmi, L, Metelli, MR, Gorini, I, Spinelli, C, Miccoli, P, Giardino, R 2002Long-term monitoring of cell-mediated immunity in disease-free breast cancer patients: a preliminary retrospective studyBiomed Pharmacother56339344PubMedGoogle Scholar
  28. 28.
    Kastelan, M, Rudolf, M, Hrsak, M, Stojkovic, R, Gamulin, S 1985Lymphocyte reactivity to mitogens and tumor sex steroid receptors in breast cancer patientsBiomed Pharmacother39442444PubMedGoogle Scholar
  29. 29.
    Levy, SM, Herberman, RB, Whiteside, T, Sanzo, K, Lee, J, Kirkwood, J 1990Perceived social support and tumor estrogen/progesterone receptor status as predictors of natural killer cell activity in breast cancer patientsPsychosom Med527385PubMedGoogle Scholar
  30. 30.
    Levy, SM, Herberman, RB, Lee, J, Whiteside, T, Kirkwood, J, McFeeley, S 1990Estrogen receptor concentration and social factors as predictors of natural killer cell activity in early-stage breast cancer patientsConfirmation of a model. Nat Immun Cell Growth Regul9313324Google Scholar
  31. 31.
    Garner, WL, Minton, JP, James, AG, Hoffmann, CC 1983Human breast cancer and impaired NK cell functionJ Surg Oncol246466PubMedGoogle Scholar
  32. 32.
    Elliott, RL, Head, JF, McCoy, JL 1994Comparison of estrogen and progesterone receptor status to lymphocyte immunity against tumor antigens in breast cancer patientsBreast Cancer Res Treat30299304PubMedGoogle Scholar
  33. 33.
    Wiltschke, C, Krainer, M, Budinsky, AC, Berger, A, Muller, C, Zeillinger, R, Speiser, P, Kubista, E, Eibl, M, Zielinski, CC 1995Reduced mitogenic stimulation of peripheral blood mononuclear cells as a prognostic parameter for the course of breast cancer: a prospective longitudinal studyBr J Cancer7112921296PubMedGoogle Scholar
  34. 34.
    Cote, RJ, Rosen, PP, Lesser, ML, Old, LJ, Osborne, MP 1991Prediction of early relapse in patients with operable breast cancer by detection of occult bone marrow micrometastasesJ Clin Oncol917491756PubMedGoogle Scholar
  35. 35.
    Mansi, JL, Easton, D, Berger, U, Gazet, JC, Ford, HT, Dearnaley, D, Coombes, RC 1991Bone marrow micrometastases in primary breast cancer: prognostic significance after 6 years’ follow-upEur J Cancer2715521555PubMedCrossRefGoogle Scholar
  36. 36.
    Harbeck, N, Untch, M, Pache, L, Eiermann, W 1994Tumour cell detection in the bone marrow of breast cancer patients at primary therapy: results of a 3-year median follow-upBr J Cancer69566571PubMedGoogle Scholar
  37. 37.
    Diel, IJ, Kaufmann, M, Costa, SD, Holle, R, Minckwitz, G, Solomayer, EF, Kaul, S, Bastert, G 1996Micrometastatic breast cancer cells in bone marrow at primary surgery: prognostic value in comparison with nodal statusJ Natl Cancer Inst8816521658PubMedGoogle Scholar
  38. 38.
    Braun, S, Pantel, K, Muller, P, Janni, W, Hepp, F, Kentenich, CR, Gastroph, S, Wischnik, A, Dimpfl, T, Kindermann, G, Riethmuller, G, Schlimok, G 2000Cytokeratin-positive cells in the bone marrow and survival of patients with stage I, II, or III breast cancerN Engl J Med342525533PubMedGoogle Scholar
  39. 39.
    Maraskovsky, E, Chen, WF, Shortman, K 1989IL-2 and IFN-gamma are two necessary lymphokines in the development of cytolytic T cellsJ Immunol14312101214PubMedGoogle Scholar
  40. 40.
    Hsieh, CS, Macatonia, SE, Tripp, CS, Wolf, SF, O’Garra, A, Murphy, KM 1993Development of TH1 CD4+ T cells through IL-12 produced by Listeria-induced macrophagesScience260547549PubMedGoogle Scholar
  41. 41.
    Manetti, R, Parronchi, P, Giudizi, MG, Piccinni, MP, Maggi, E, Trinchieri, G, Romagnani, S 1993Natural killer cell stimulatory factor (interleukin 12 [IL-12]) induces T helper type 1 (Th1)-specific immune responses and inhibits the development of IL-4-producing Th cellsJ Exp Med17711991204PubMedGoogle Scholar
  42. 42.
    Liyanage, UK, Moore, TT, Joo, HG, Tanaka, Y, Herrmann, V, Doherty, G, Drebin, JA, Strasberg, SM, Eberlein, TJ, Goedegebuure, PS, Linehan, DC 2002Prevalence of regulatory T cells is increased in peripheral blood and tumor microenvironment of patients with pancreas or breast adenocarcinomaJ Immunol16927562761PubMedGoogle Scholar
  43. 43.
    Wolf, AM, Wolf, D, Steurer, M, Gastl, G, Gunsilius, E, Grubeck-Loebenstein, B 2003Increase of regulatory T cells in the peripheral blood of cancer patientsClin Cancer Res9606612PubMedGoogle Scholar
  44. 44.
    Burford-Mason, A, Gyte, GM, Watkins, SM 1989Phytohaemagglutinin responsiveness of peripheral lymphocytes and survival in patients with primary breast cancerBreast Cancer Res Treat13243250PubMedGoogle Scholar
  45. 45.
    Wiltschke, C, Krainer, M, Budinsky, AC, Berger, A, Muller, C, Zeillinger, R, Speiser, P, Kubista, E, Eibl, M, Zielinski, CC 1995Reduced mitogenic stimulation of peripheral blood mononuclear cells as a prognostic parameter for the course of breast cancer: a prospective longitudinal studyBr J Cancer7112921296PubMedGoogle Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • Michael J. Campbell
    • 1
  • Janet Scott
    • 2
  • Holden T. Maecker
    • 3
  • John W. Park
    • 2
  • Laura J. Esserman
    • 1
  1. 1.Department of SurgeryUniversity of California, UCSF/Mt. Zion Med CtrSan Francisco
  2. 2.Department of MedicineUniversity of CaliforniaSan FranciscoUSA
  3. 3.BD BiosciencesSan JoseUSA

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