Cancer Immunology, Immunotherapy

, Volume 57, Issue 2, pp 271–280 | Cite as

Adoptive transfer of autologous, HER2-specific, cytotoxic T lymphocytes for the treatment of HER2-overexpressing breast cancer

  • Helga Bernhard
  • Julia Neudorfer
  • Kerstin Gebhard
  • Heinke Conrad
  • Christine Hermann
  • Jörg Nährig
  • Falko Fend
  • Wolfgang Weber
  • Dirk H. Busch
  • Christian Peschel
Short Communication


The human epidermal growth factor receptor 2 (HER2) has been targeted as a breast cancer-associated antigen by immunotherapeutical approaches based on HER2-directed monoclonal antibodies and cancer vaccines. We describe the adoptive transfer of autologous HER2-specific T-lymphocyte clones to a patient with metastatic HER2-overexpressing breast cancer. The HLA/multimer-based monitoring of the transferred T lymphocytes revealed that the T cells rapidly disappeared from the peripheral blood. The imaging studies indicated that the T cells accumulated in the bone marrow (BM) and migrated to the liver, but were unable to penetrate into the solid metastases. The disseminated tumor cells in the BM disappeared after the completion of adoptive T-cell therapy. This study suggests the therapeutic potential for HER2-specific T cells for eliminating disseminated HER2-positive tumor cells and proposes the combination of T cell-based therapies with strategies targeting the tumor stroma to improve T-cell infiltration into solid tumors.


Tumor immunity Human Cytotoxic T cells Antigens/peptides/epitopes MHC 



Human epidermal growth factor receptor 2




Single photon emission computed tomography


Magnetic resonance tomography


[18F] Fluorodeoxyglucose positron emission tomography


Mononuclear cell



We thank the patients for taking part in this clinical trial; Burkhard Schmidt, Evelyn Schulz and Matthias Schiemann for excellent technical assistance; Peter Schmidkonz for expert clinical care; and Wendy Batten for helpful discussion and critical reading of the manuscript. This work was supported by the Research Council of Germany grant SFB 456 (to H.B. and D.H.B.) and the Wilhelm Sander-Stiftung grant 2000.017.3 (to H.B.).

Supplementary material

262_2007_355_MOESM1_ESM.doc (40 kb)
ESM1 (DOC 40.5 kb)


  1. 1.
    Qin Z, Blankenstein T (2004) A cancer immunosurveillance controversy. Nat Immunol 5:3–4PubMedCrossRefGoogle Scholar
  2. 2.
    Lee PP, Yee C, Savage PA, Fong L, Brockstedt D, Weber JS, Johnson D, Swetter S, Thompson J, Greenberg PD, Roederer M, Davis MM (1999) Characterization of circulating T cells specific for tumor-associated antigens in melanoma patients. Nat Med 5:677–685PubMedCrossRefGoogle Scholar
  3. 3.
    Jäger E, Nagata Y, Gnjatic S, Wada H, Stockert E, Karbach J, Dunbar PR, Lee SY, Jungblut A, Jäger D, Arand M, Ritter G, Cerundolo V, Dupont B, Chen Y-T, Old LJ, Knuth A (2000) Monitoring CD8 T cell responses to NY-ESO-1: correlation of humoral and cellular immune responses. Proc Natl Acad Sci USA 97:4760–4765PubMedCrossRefGoogle Scholar
  4. 4.
    Willimsky G, Blankenstein T (2005) Sporadic immunogenic tumours avoid destruction by inducing T-cell tolerance. Nature 437:141–146PubMedCrossRefGoogle Scholar
  5. 5.
    Zhang L, Conejo-Garcia JR, Katsaros D, Gimotty PA, Massobrio M, Regnani G, Makrigiannakis A, Gray H, Schlienger K, Liebman MN, Rubin SC, Coukos G (2003) Intratumoral T cells, recurrence, and survival in epithelial ovarian cancer. N Engl J Med 348:203–213PubMedCrossRefGoogle Scholar
  6. 6.
    Öhlén C, Kalos M, Hong DJ, Shur AC, Greenberg PD (2001) Expression of a tolerizing tumor antigen in peripheral tissue does not preclude recovery of high-affinity CD8+ T cells or CTL immunotherapy of tumors expressing the antigen. J Immunol 166:2863–2870PubMedGoogle Scholar
  7. 7.
    Teague RM, Sather BD, Sacks JA, Huang MZ, Dossett ML, Morimoto J, Tan S, Sutton SE, Cooke MP, Öhlén C, Greenberg PD (2006) Interleukin-15 rescues tolerant CD8+ T cells for use in adoptive immunotherapy of established tumors. Nat Med 12:335–341PubMedCrossRefGoogle Scholar
  8. 8.
    Bollard CM, Aguilar L, Straathof KC, Gahn B, Huls MH, Rousseau A, Sixbey J, Gresik MV, Carrum G, Hudson M, Dilloo D, Gee A, Brenner MK, Rooney CM, Heslop HE (2004) Cytotoxic T lymphocyte therapy for Epstein–Barr virus Hodgkin’s disease. J Exp Med 200:1623–1633PubMedCrossRefGoogle Scholar
  9. 9.
    Yee C, Thompson JA, Byrd D, Riddell SR, Roche P, Celis E, Greenberg PD (2002) Adoptive T cell therapy using antigen-specific CD8+ T cell clones for the treatment of patients with metastatic melanoma: In vivo persistence, migration, and antitumor effect of transferred T cells. PNAS 99:16168–16173PubMedCrossRefGoogle Scholar
  10. 10.
    Dudley ME, Wunderlich JR, Robbins PF, Yang JC, Hwu P, Schwartzentruber DJ, Topalian SL, Sherry R, Restifo NP, Hubicki AM, Robinson MR, Raffeld M, Duray P, Seipp CA, Rogers-Freezer L, Morton KE, Mavroukakis SA, White DE, Rosenberg SA (2002) Cancer regression and autoimmunity in patients after clonal repopulation with antitumor lymphocytes. Science 298:850–854PubMedCrossRefGoogle Scholar
  11. 11.
    Meidenbauer N, Marienhagen J, Laumer M, Vogl S, Heymann J, Andreesen R, Mackensen A (2003) Survival and tumor localization of adoptively transferred melan-A-specific T cells in melanoma patients. J Immunol 170:2161–2169PubMedGoogle Scholar
  12. 12.
    Disis ML, Gooley TA, Rinn K, Davis D, Piepkorn M, Cheever MA, Knutson KL, Schiffman K (2002) Generation of T-cell immunity to the HER-2/neu protein after active immunization with HER-2/neu peptide-based vaccines. J Clin Oncol 20:2624–2632PubMedCrossRefGoogle Scholar
  13. 13.
    Blankenstein T (2005) The role of tumor stroma in the interaction between tumor and immune system. Curr Opin Immunol 17:180–186PubMedCrossRefGoogle Scholar
  14. 14.
    Knabel M, Franz TJ, Schiemann M, Wulf A, Villmow B, Schmidt B, Bernhard H, Wagner H, Busch DH (2002) Reversible MHC multimer staining for functional isolation of T-cell populations and effective adoptive transfer. Nat Med 8:631–637PubMedCrossRefGoogle Scholar
  15. 15.
    Meyer zum Büschenfelde C, Metzger J, Hermann C, Nicklisch N, Peschel C, Bernhard H (2001) The generation of both T killer and T helper cell clones specific for the tumor-associated antigen HER2 using retrovirally transduced dendritic cells. J Immunol 167:1712–1719Google Scholar
  16. 16.
    Walker BD, Flexner C, Birch-Limberger K, Fisher L, Paradis TJ, Aldovini A, Young R, Moss B, Schooley RT (1989) Long-term culture and fine specificity of human cytotoxic T-lymphocyte clones reactive with human immunodeficiency virus type 1. Proc Natl Acad Sci USA 86:9514–9518PubMedCrossRefGoogle Scholar
  17. 17.
    Braun S, Vogl FD, Naume B, Janni W, Osborne MP, Coombes C, Schlimok G, Diel IJ, Gerber B, Gebauer G, Pierga J-Y, Marth C, Oruzio D, Wiedswang G, Solomayer E-F, Kundt G, Strobl B, Fehm T, Wong GYC, Bliss J, Vincent-Salomon A, Pantel K (2005) A pooled analysis of bone marrow micrometastasis in breast cancer. N Engl J Med 353:793–802PubMedCrossRefGoogle Scholar
  18. 18.
    Fisk B, Blevins TL, Wharton JT, Ioannides CG (1995) Identification of an immunodominant peptide of HER-2/neu protooncogene recognized by ovarian tumor-specific cytotoxic T lymphocyte lines. J Exp Med 181:2109–2117PubMedCrossRefGoogle Scholar
  19. 19.
    Neudorfer J, Schmidt B, Huster KM, Anderl F, Schiemann M, Holzapfel G, Schmidt T, Germeroth L, Wagner H, Peschel C, Busch DH, Bernhard H (2007) Reversible HLA multimers (Streptamers) for the isolation of human cytotoxic T lymphocytes functionally active against tumor- and virus-derived antigens. J Immunol Methods 320:119–131PubMedCrossRefGoogle Scholar
  20. 20.
    Rongcun Y, Salazar-Onfray F, Charo J, Malmberg K-J, Evrin K, Maes H, Kono K, Hising C, Petersson M, Larsson O, Lan L, Appella E, Sette A, Celis E, Kiesling R (1999) Identification of new HER2/neu-derived peptide epitopes that can elicit specific CTL against autologous and allogeneic carcinomas and melanomas. J Immunol 163:1037–1044PubMedGoogle Scholar
  21. 21.
    Zaks TZ, Rosenberg SA (1998) Immunization with a peptide epitope (p369–377) from HER-2/neu leads to peptide-specific cytotoxic T lymphocytes that fail to recognize HER-2/neu+ tumors. Cancer Res 58:4902–4908PubMedGoogle Scholar
  22. 22.
    Herrmann F, Lehr H-A, Drexler I, Sutter G, Hengstler J, Wollscheid U, Seliger B (2004) HER-2/neu-mediated regulation of components of the MHC class I antigen-processing pathway. Cancer Res 64:215–220PubMedCrossRefGoogle Scholar
  23. 23.
    Castilleja A, Carter D, Efferson CL, Ward NE, Kawano K, Fisk B, Kudelka AP, Gershenson DM, Muarray JL, O’Brian CA, Ioannides CG (2002) Induction of tumor-reactive CTL by c-side chain variants of the CTL epitope HER-2/neu protooncogene (369–377) selected by molecular modeling of the peptide: HLA-A2 complex. J Immunol 169:3545–3554PubMedGoogle Scholar
  24. 24.
    Vertuani S, Sette A, Sidney J, Southwood S, Fikes J, Keogh E, Lindencrona JA, Ishioka G, Levitskaya J, Kiessling R (2004) Improved immunogenicity of an immunodominant epitope of the Her-2/neu protooncogene by alterations of MHC contact residues. J Immunol 172:3501–3508PubMedGoogle Scholar
  25. 25.
    Lustgarten J, Dominguez AL, Pinilla C (2006) Identification of cross-reactive peptides using combinatorial libraries circumvents tolerance against Her-2/neu-immunodominant epitope. J Immunol 176:1796–1805PubMedGoogle Scholar
  26. 26.
    Müller MR, Grünebach F, Nencioni A, Brossart P (2003) Transfection of dendritic cells with RNA induces CD4- and CD8-mediated T cell immunity against breast carcinomas and reveals the immunodominance of presented T cell epitopes. J Immunol 170:5892–5896PubMedGoogle Scholar
  27. 27.
    Ganss R, Ryschich E, Klar E, Arnold B, Hämmerling GJ (2002) Combination of T-cell therapy and trigger of inflammation induces remodeling of the vasculature and tumor eradication. Cancer Res 62:1462–1470PubMedGoogle Scholar
  28. 28.
    Padera TP, Stoll BR, Tooredman JB, Capen D, diTomaso E, Jain RK (2004) Pathology: cancer cells compress intratumour vessels. Nature 427:695PubMedCrossRefGoogle Scholar
  29. 29.
    Garbi N, Arnold B, Gordon S, Hämmerling GJ, Ganss R (2004) CpG motifs as proinflammatory factors render autochthonous tumors permissive for infiltration and destruction. J Immunol 172:5861–5869PubMedGoogle Scholar
  30. 30.
    Feuerer M, Beckhove P, Bai L, Solomayer E-F, Bastert G, Diel IJ, Pedain C, Oberniedermayr M, Schirrmacher V, Umansky V (2001) Therapy of human tumors in NOD/SCID mice with patient-derived reactivated memory T cells from bone marrow. Nature Med 7:452–458PubMedCrossRefGoogle Scholar
  31. 31.
    Carson WE, Parihar R, Lindemann MJ, Personeni N, Dierksheide J, Meropol NJ, Baselga J, Caligiuri MA (2001) Interleukin-2 enhances the natural killer cell response to Herceptin-coated Her2/neu-positive breast cancer cells. Eur J Immunol 31:3016–3025PubMedCrossRefGoogle Scholar
  32. 32.
    Fleming GF, Meropol NJ, Rosner GL, Hollis DR, Carson WE, Caligiuri M, Mortimer J, Tkaczuk K, Parihar R, Schilsky RL, Ratain MJ (2002) A phase I trial of escalating doses of trastuzumab combined with daily subcutaneous interleukin 2: report of Cancer and Leukemia Group B 9661. Clin Cancer Res 8:3718–3727PubMedGoogle Scholar
  33. 33.
    Meyer zum Büschenfelde C, Hermann C, Schmidt B, Peschel C, Bernhard H (2002) Antihuman epidermal growth factor receptor 2 (HER2) monoclonal antibody trastuzumab enhances cytolytic activity of class I-restricted HER2-specific T lymphocytes against HER2-overexpressing tumor cells. Cancer Research 62:2244–2247Google Scholar
  34. 34.
    Gattinoni L, Klebanoff CA, Palmer DC, Wrzesinski C, Kerstann K, Yu Z, Finkelstein SE, Theroret MR, Rosenberg SA, Restifo NP (2005) Acquisition of full effector function in vitro paradoxically impairs the in vivo antitumor efficacy of adoptively transferred CD8+ T cells. J Clin Invest 115:1616–1626PubMedCrossRefGoogle Scholar
  35. 35.
    Huster KM, Busch V, Schiemann M, Linkemann K, Kerksiek KM, Wagner H, Busch DH (2004) Selective expression of IL-7 receptor on memory T cells identifies early CD40L-dependent generation of distinct CD8+ memory T cell subsets. PNAS 101:5610–5615PubMedCrossRefGoogle Scholar
  36. 36.
    Robbins PF, Dudley ME, Wunderlich J, El-Gamil M, Li YF, Zhou J, Huang J, Powell DJ, Rosenberg SA (2004) Persistence of transferred lymphocyte clonotypes correlates with cancer regression in patients receiving cell transfer therapy. J Immunol 173:7125–7130PubMedGoogle Scholar
  37. 37.
    Zhou J, Dudley ME, Rosenberg SA, Robbins PF (2004) Selective growth, in vitro and in vivo, of individual T cell clones from tumor-infiltrating lymphocytes obtained from patients with melanoma. J Immunol 173:7622–7629PubMedGoogle Scholar
  38. 38.
    Zhou J, Shen X, Huang J, Hodes RJ, Rosenberg SA, Robbins PF (2005) Telomere length of transferred lymphocytes correlates with in vivo persistence and tumor regression in melanoma patients receiving cell transfer therapy. J Immunol 175:7046–7052PubMedGoogle Scholar
  39. 39.
    Berger C, Huang M-L, Gough M, Greenberg PD, Riddell SR, Kiem H-P (2000) Nonmyeloablative immunosuppressive regimen prolongs in vivo persistence of gene-modified autologous T cells in a nonhuman primate model. J Virol 75:799–808CrossRefGoogle Scholar
  40. 40.
    Dannull J, Su Z, Rizzieri D, Yang BK, Coleman D, Yancey D, Zhang A, Dahm P, Chao N, Gilboa E, Vieweg J (2005) Enhancement of vaccine-mediated antitumor immunity in cancer patients after depletion of regulatory T cells. J Clin Invest 115:3623–3633PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Helga Bernhard
    • 1
    • 5
    • 6
  • Julia Neudorfer
    • 1
  • Kerstin Gebhard
    • 1
    • 5
  • Heinke Conrad
    • 1
  • Christine Hermann
    • 1
  • Jörg Nährig
    • 2
  • Falko Fend
    • 2
  • Wolfgang Weber
    • 3
  • Dirk H. Busch
    • 4
    • 5
  • Christian Peschel
    • 1
  1. 1.Department of Hematology/OncologyTechnical University of MunichMunichGermany
  2. 2.Department of PathologyTechnical University of MunichMunichGermany
  3. 3.Department of Nuclear MedicineTechnical University of MunichMunichGermany
  4. 4.Department of Microbiology, Immunology and HygieneTechnical University of MunichMunichGermany
  5. 5.Clinical Cooperation Group ‘Antigen-Specific Immunotherapy’GSF, Institute of Health and Environment Neuherberg and Technical University of MunichNeuherbergGermany
  6. 6.3rd Medical DepartmentTechnical University of MunichMunichGermany

Personalised recommendations