Annals of Surgical Oncology

, Volume 12, Issue 7, pp 517–525 | Cite as

A Tat Fusion Protein–Based Tumor Vaccine for Breast Cancer

  • Carsten T. Viehl
  • Michelle Becker-Hapak
  • Jason S. Lewis
  • Yoshiyuki Tanaka
  • Udaya K. Liyanage
  • David C. Linehan
  • Timothy J. Eberlein
  • Peter S. Goedegebuure



We recently reported that dendritic cells (DCs) transduced with a fusion protein between Her2/neu and the protein transduction domain Tat (DC-Tat-extracellular domain [ECD]) induced Her2/neu-specific CD8+ T cells in vitro. This study tested the in vivo efficacy of DC-Tat-ECD in a murine breast cancer model.


FVB/N mice received one or two weekly intraperitoneal immunizations with syngeneic DC-Tat-ECD followed by a tumor challenge with syngeneic neu+ breast cancer cells, and tumor development was monitored. To test for Her2/neu specificity, CD4+ and CD8+ cells were isolated through magnetic bead separation and analyzed for specific interferon γ release.


Intraperitoneally injected DCs migrated to secondary lymphoid organs, as evidenced by small-animal positron emission tomography studies. Immunized mice developed palpable tumors significantly later than control mice injected with DC-Tat-empty (P = .001 and P < .05 for two immunizations and for one immunization, respectively) or mice that received no DCs (P = .001 and P < .05). Similarly, immunized mice had smaller resulting tumors than mice injected with DC-Tat-empty (P < .05 and P < .01) or untreated mice (P < .001 and P < .001). Significantly more tumor-specific CD8+ splenocytes were found in twice-immunized mice than in untreated animals (P < .001). Similarly, a T-helper type 1 CD4+ T-cell response was observed.


Protein-transduced DCs may be effective vaccines for the treatment of cancer.


Tat fusion protein Her2/neu Cancer vaccine Breast cancer Animal study 


  1. 1.
    Street, SE, Hayakawa, Y, Zhan, Y,  et al. 2004Innate immune surveillance of spontaneous B cell lymphomas by natural killer cells and γδ T cellsJ Exp Med19987984PubMedGoogle Scholar
  2. 2.
    Shankaran, V, Ikeda, H, Bruce, AT,  et al. 2001IFNgamma and lymphocytes prevent primary tumour development and shape tumour immunogenicityNature410110711CrossRefPubMedGoogle Scholar
  3. 3.
    Ribas, A, Butterfield, LH, Glaspy, JA, Economou, JS 2003Current developments in cancer vaccines, cellular immunotherapyJ Clin Oncol21241532CrossRefPubMedGoogle Scholar
  4. 4.
    Tanaka, Y, Dowdy, SF, Linehan, DC, Eberlein, TJ, Goedegebuure, PS 2003Induction of antigen-specific CTL by recombinant HIV trans-activating fusion protein-pulsed human monocyte-derived dendritic cellsJ Immunol17012918PubMedGoogle Scholar
  5. 5.
    Schwarze, SR, Hruska, KA, Dowdy, SF 2000Protein transduction: unrestricted delivery into all cells?Trends Cell Biol 102905Google Scholar
  6. 6.
    Becker-Hapak, M, McAllister, SS, Dowdy, SF 2001TAT-mediated protein transduction into mammalian cellsMethods2424756PubMedGoogle Scholar
  7. 7.
    Wadia, JS, Dowdy, SF 2002Protein transduction technologyCurr Opin Biotechnol13526PubMedGoogle Scholar
  8. 8.
    Fawell, S, Seery, J, Daikh, Y,  et al. 1994Tat-mediated delivery of heterologous proteins into cellsProc Natl Acad Sci U S A916648PubMedGoogle Scholar
  9. 9.
    Kim, DT, Mitchell, DJ, Brockstedt, DG,  et al. 1997Introduction of soluble proteins into the MHC class I pathway by conjugation to an HIV tat peptideJ Immunol15916668PubMedGoogle Scholar
  10. 10.
    Lu, J, Wettstein, PJ, Higashimoto, Y, Appella, E, Celis, E 2001TAP-independent presentation of CTL epitopes by Trojan antigensJ Immunol166706371PubMedGoogle Scholar
  11. 11.
    Shibagaki, N, Udey, MC 2002Dendritic cells transduced with protein antigens induce cytotoxic lymphocytes and elicit antitumor immunityJ Immunol1682393401PubMedGoogle Scholar
  12. 12.
    Wang, HY, Fu, T, Wang, G,  et al. 2002Induction of CD4(+) T cell-dependent antitumor immunity by TAT-mediated tumor antigen delivery into dendritic cellsJ Clin Invest109146370CrossRefPubMedGoogle Scholar
  13. 13.
    Shibagaki, N, Udey, MC 2003Dendritic cells transduced with TAT protein transduction domain-containing tyrosinase-related protein 2 vaccinate against murine melanomaEur J Immunol3385060CrossRefPubMedGoogle Scholar
  14. 14.
    Viehl CT, Tanaka Y, Chen T et al. Tat mammaglobin fusion protein transduced dendritic cells stimulate mammoglobin-specific CD4 and CD8 T cells Breast Cancer Res Treat 2005, DOI: 10.1007/s10549-005-0450-4 Google Scholar
  15. 15.
    Yarden, Y 2001Biology of HER2 and its importance in breast cancerOncology61113CrossRefGoogle Scholar
  16. 16.
    Slamon, DJ, Clark, GM, Wong, SG, Levin, WJ, Ullrich, A, McGuire, WL 1987Human breast cancer: correlation of relapse, survival with amplification of the HER-2/neu oncogeneScience23517782PubMedGoogle Scholar
  17. 17.
    Disis, ML, Calenoff, E, McLaughlin, G,  et al. 1994Existent T-cell and antibody immunity to HER-2/neu protein in patients with breast cancerCancer Res541620PubMedGoogle Scholar
  18. 18.
    Peoples, GE, Schoof, DD, Andrews, JV, Goedegebuure, PS, Eberlein, TJ 1993T-cell recognition of ovarian cancerSurgery11422734PubMedGoogle Scholar
  19. 19.
    Yoshino, I, Goedegebuure, PS, Peoples, GE,  et al. 1994HER2/neu-derived peptides are shared antigens among human non-small cell lung cancer and ovarian cancerCancer Res54338790Google Scholar
  20. 20.
    Peoples, GE, Goedegebuure, PS, Smith, R, Linehan, DC, Yoshino, I, Eberlein, TJ 1995Breast and ovarian cancer-specific cytotoxic T lymphocytes recognize the same HER2/neu-derived peptideProc Natl Acad Sci U S A924326PubMedGoogle Scholar
  21. 21.
    Tanaka, Y, Amos, KD, Joo, HG, Eberlein, TJ, Goedegebuure, PS 2001Modification of the HER2/NEU-derived tumor antigen GP2 improves induction of GP2-reactive cytotoxic T lymphocytesInt J Cancer945404Google Scholar
  22. 22.
    Fisk, B, Blevins, TL, Wharton, JT, Ioannides, CG 1995Identification of an immunodominant peptide of HER-2/neu protooncogene recognized by ovarian tumor-specific cytotoxic T lymphocyte linesJ Exp Med181210917PubMedGoogle Scholar
  23. 23.
    Kobayashi, H, Wood, M, Song, Y, Appella, E, Celis, E 2000Defining promiscuous MHC class II helper T-cell epitopes for the HER2/neu tumor antigenCancer Res60522836Google Scholar
  24. 24.
    Reilly, RT, Gottlieb, MB, Ercolini, AM,  et al. 2000HER-2/neu is a tumor rejection target in tolerized HER-2/neu transgenic miceCancer Res60356976Google Scholar
  25. 25.
    Yoshino, I, Peoples, GE, Goedegebuure, PS, Maziarz, R, Eberlein, TJ 1994Association of HER2/neu expression with sensitivity to tumor-specific CTL in human ovarian cancerJ Immunol1522393400PubMedGoogle Scholar
  26. 26.
    Nagahara, H, Vocero-Akbani, AM, Snyder, EL,  et al. 1998Transduction of full-length TAT fusion proteins into mammalian cells: TAT-p27Kip1 induces cell migrationNat Med4144952CrossRefPubMedGoogle Scholar
  27. 27.
    McCarthy, DW, Shefer, RE, Klinkowstein, RE,  et al. 1997Efficient production of high specific activity 64Cu using a biomedical cyclotronNucl Med Biol243543CrossRefPubMedGoogle Scholar
  28. 28.
    McCarthy, DW, Bass, LA, Cutler, PD,  et al. 1999High purity production and potential applications of copper-60 and copper-61Nucl Med Biol263518CrossRefPubMedGoogle Scholar
  29. 29.
    Fujibayashi, Y, Taniuchi, H, Yonekura, Y, Ohtani, H, Konishi, J 1997Yokoyama Copper-62-ATSM: a new hypoxia imaging agent with high membrane permeability and low redox potentialJ Nucl Med38115560PubMedGoogle Scholar
  30. 30.
    Young, H, Carnochan, P, Zweit, J, Babich, J, Cherry, S, Ott, R 1994Evaluation of copper(II)-pyruvaldehyde bis (N-4-methylthiosemicarbazone) for tissue blood flow measurement using a trapped tracer modelEur J Nucl Med2133641PubMedGoogle Scholar
  31. 31.
    Defrise, M, Kinahan, PE, Townsend, DW, Michel, C, Sibomana, M, Newport, DF 1997Exact and approximate rebinning algorithms for 3-D PET dataIEEE Trans Med Imaging1614558PubMedGoogle Scholar
  32. 32.
    Janik, P, Briand, P, Hartmann, NR 1975The effect of estrone-progesterone treatment on cell proliferation kinetics of hormone-dependent GR mouse mammary tumorsCancer Res353698704Google Scholar
  33. 33.
    Banchereau, J, Steinman, RM 1998Dendritic cells and the control of immunityNature39224552PubMedGoogle Scholar
  34. 34.
    Frankel, AD, Pabo, CO 1988Cellular uptake of the tat protein from human immunodeficiency virusCell55118993CrossRefPubMedGoogle Scholar
  35. 35.
    Green, M, Loewenstein, PM 1988Autonomous functional domains of chemically synthesized human immunodeficiency virus tat trans-activator proteinCell55117988PubMedGoogle Scholar
  36. 36.
    Wang, RF, Wang, HY 2002Enhancement of antitumor immunity by prolonging antigen presentation on dendritic cellsNat Biotechnol2014954CrossRefPubMedGoogle Scholar
  37. 37.
    Lu, J, Higashimoto, Y, Appella, E, Celis, E 2004Multiepitope Trojan antigen peptide vaccines for the induction of antitumor CTL and Th immune responsesJ Immunol172457582PubMedGoogle Scholar
  38. 38.
    Viehl, CT, Liyanage, UK, Moore, TT,  et al. 2004Depletion of regulatory T cells promotes a tumor-specific immune responseAnn Surg Oncol11S69Google Scholar
  39. 39.
    Wei, WZ, Shi, WP, Galy, A,  et al. 1999Protection against mammary tumor growth by vaccination with full-length, modified human ErbB-2 DNAInt J Cancer8174854Google Scholar
  40. 40.
    Pilon, SA, Piechocki, MP, Wei, WZ 2001Vaccination with cytoplasmic ErbB-2 DNA protects mice from mammary tumor growth without anti-ErbB-2 antibodyJ Immunol16732016PubMedGoogle Scholar
  41. 41.
    Piechocki, MP, Pilon, SA, Wei, WZ 2001Complementary antitumor immunity induced by plasmid DNA encoding secreted and cytoplasmic human ErbB-2J Immunol167336774PubMedGoogle Scholar
  42. 42.
    Piechocki, MP, Ho, YS, Pilon, S, Wei, WZ 2003Human ErbB-2 (Her-2) transgenic mice: a model system for testing Her-2 based vaccinesJ Immunol171578794PubMedGoogle Scholar
  43. 43.
    Baselga, J, Tripathy, D, Mendelsohn, J,  et al. 1996Phase II study of weekly intravenous recombinant humanized anti-p185HER2 monoclonal antibody in patients with HER2/neu-overexpressing metastatic breast cancerJ Clin Oncol1473744PubMedGoogle Scholar
  44. 44.
    Slamon, DJ, Leyland-Jones, B, Shak, S,  et al. 2001Use of chemotherapy plus a monoclonal antibody against HER2 for metastatic breast cancer that overexpresses HER2N Engl J Med34478392PubMedGoogle Scholar
  45. 45.
    Ko, BK, Kawano, K, Murray, JL,  et al. 2003Clinical studies of vaccines targeting breast cancerClin Cancer Res9322234PubMedGoogle Scholar
  46. 46.
    Rosenberg, SA 2004Development of effective immunotherapy for the treatment of patients with cancerJ Am Coll Surg19868596CrossRefPubMedGoogle Scholar

Copyright information

© The Society of Surgical Oncology, Inc. 2005

Authors and Affiliations

  • Carsten T. Viehl
    • 1
    • 2
  • Michelle Becker-Hapak
    • 3
  • Jason S. Lewis
    • 4
    • 5
  • Yoshiyuki Tanaka
    • 1
  • Udaya K. Liyanage
    • 1
  • David C. Linehan
    • 1
    • 5
  • Timothy J. Eberlein
    • 1
    • 5
  • Peter S. Goedegebuure
    • 1
    • 5
  1. 1.Department of SurgeryWashington University School of MedicineSt. Louis
  2. 2.Department of Surgery, Divisions of General Surgery and Surgical ResearchUniversity of BaselSwitzerland
  3. 3.Division of OncologyAlvin J. Siteman Cancer Center, Washington University School of MedicineSt. Louis
  4. 4.Mallinckrodt Institute of RadiologyWashington University School of MedicineSt. Louis
  5. 5.Alvin J. Siteman Cancer CenterWashington University School of MedicineSt. Louis

Personalised recommendations