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A phage display selected Fab fragment with MHC class I-restricted specificity for MAGE-A1 allows for retargeting of primary human T lymphocytes

Gene Therapy volume 8, pages 1601–1608 (2001)Cite this article

Abstract

The clinical benefit of adoptive transfer of MHC-restricted cytotoxic T lymphocytes(CTL) for the treatment of cancer is hampered by the low success rate to generate antitumor CTLs. To bypass the need for tumor-specific CTL, we developed a strategy that allows for grafting of human T lymphocytes with MHC-restricted antigen specificity using in vitro selected human Fab fragments fused to the Fc(ɛ)RI-γ signaling molecule. Retroviral introduction of a Fab-based chimeric receptor specific for MAGE-A1/HLA-A1 into primary human T lymphocytes resulted in binding of relevant peptide/MHC complexes. Transduced T lymphocytes responded to native MAGE-A1/HLA-A1POS target cells by specific cytokine production and cytolysis. Therefore, peptide/MHC-specific Fab fragments represent new alternatives to TCR to confer human T lymphocytes with tumor specificity, which provides a promising rationale for developing immunogene therapies. Gene Therapy (2001) 8, 000-000.

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References

  1. Meuer SC . Identification of the receptor for antigen and major histocompatibility complex on human inducer T lymphocytes Science 1983 4629: 1239–1242

    Article  Google Scholar 

  2. Borst JA . T-cell receptor gamma/CD3 complex found on cloned functional lymphocytes Nature 1987 6106: 683–688

    Article  Google Scholar 

  3. Davis MM, Bjorkman PJ . T-cell antigen receptor genes and T-cell recognition Nature 1988 334: 395–402

    Article  CAS  PubMed  Google Scholar 

  4. Klausner RD, Lippincott-Schwartz J, Bonifacio JS . The T cell antigen receptor: insight into organelle biology Ann Rev Cell Biol 1990 6: 403–431

    Article  CAS  PubMed  Google Scholar 

  5. Riddell SR . Restoration of viral immunity in immunodeficient humans by adoptive transfer of T cell clones Science 1992 257: 238–241

    Article  CAS  PubMed  Google Scholar 

  6. Rosenberg SA . Treatment of patients with metastatic melanoma with autologous tumor-infiltrating lymphocytes and interleukin 2 J Natl Cancer Inst 1994 86: 1159–1166

    Article  CAS  PubMed  Google Scholar 

  7. Van der Bruggen P . A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma Science 1991 254: 1643–1647

    Article  CAS  PubMed  Google Scholar 

  8. Boon T, van der Bruggen P . Human tumor antigens recognized by T lymphocytes J Exp Med 1996 183: 725–729

    Article  CAS  PubMed  Google Scholar 

  9. Heemels MT, Ploegh H . Generation, translocation, and presentation of MHC class I-restricted peptides Annu Rev Biochem 1995 64: 463–491

    Article  CAS  PubMed  Google Scholar 

  10. Madden DR, Gorga JC, Strominger JL, Wiley DC . The structure of HLA-B27 reveals nonamer self-peptides bound in an extended conformation Nature 1995 353: 321–325

    Article  Google Scholar 

  11. Rammensee HG, Falk K, Rotzschke O . Peptides naturally presented by MHC class I molecules Annu Rev Immunol 1993 11: 213–244

    Article  CAS  PubMed  Google Scholar 

  12. Garboczi DN . Structure of the complex between human T-cell receptor, viral peptide and HLA-A2 Nature 1996 384: 134–141

    Article  CAS  PubMed  Google Scholar 

  13. Rosenberg SA . Cancer vaccines based on the identification of genes encoding cancer regression antigens Immunol Today 1997 4: 75–182

    Google Scholar 

  14. Rosenberg SA . Immunologic and therapeutic evaluation of a synthetic peptide vaccine for the treatment of patients with metastatic melanoma Nat Med 1998 4: 321–327

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Nestle FO . Vaccination of melanoma patients with peptide- or tumor lysate-pulsed dendritic cells Nat Med 1998 4: 328–332

    Article  CAS  PubMed  Google Scholar 

  16. Yee C . Isolation of high avidity melanoma-reactive CTL from heterogeneous populations using peptide-MHC tetramers J Immunol 1999 162: 2227–2234

    CAS  PubMed  Google Scholar 

  17. Zaks TZ, Rosenberg ST . Immunization with a peptide epitope from HER-2/neu leads to peptide-specific cytotoxic T lymphocytes that fail to recognize HER-2/neu+ tumors Cancer Res 1998 58: 4902–4908

    CAS  PubMed  Google Scholar 

  18. Rivoltini L . Induction of tumor-reactive CTL from peripheral blood and tumor-infiltrating lymphocytes of melanoma patients by in vitro stimulation with an immunodominant peptide of the human melanoma antigen MART-1 J Immunol 1995 54: 2257–2265

    Google Scholar 

  19. Bolhuis RLH, Gratama JW . Genetic re-targeting of T lymphocyte specificity Gene Therapy 1998 5: 1153–1155

    Article  CAS  PubMed  Google Scholar 

  20. Bolhuis RLH, Willemsen RA, Gratama JW . Clinical applications of redirected cytotoxicity Cytotoxic Cells: Basic Mechanisms and Medical Applications Lippincott Williams & Wilkins 2000 423–440

  21. Hwu P . Lysis of ovarian cancer cells by human lymphocytes redirected with a chimeric gene composed of an antibody variable region and the Fc receptor gamma chain J Exp Med 1993 178: 361–366

    Article  CAS  PubMed  Google Scholar 

  22. Weijtens MEM . Single chain Ig/γ gene-redirected human T lymphocytes produce cytokines, specifically lyse tumor cells, and recycle lytic capacity J Immunol 1996 157: 836–843

    CAS  PubMed  Google Scholar 

  23. Weijtens MEM, Willemsen RA, Hart EH, Bolhuis RLH . A retroviral vector system ‘STITCH’ in combination with an optimized single chain antibody chimeric receptor gene structure allows efficient gene transduction and expression in human T lymphocytes Gene Therapy 1998 9: 1195–1203

    Article  Google Scholar 

  24. Roberts MR . Targeting of human immunodeficiency virus-infected cells by CD8+ T lymphocytes armed with universal T-cell receptors Blood 1994 9: 2878–2889

    Google Scholar 

  25. Weijtens MEM, Willemsen RA, Van Krimpen BA, Bolhuis RLH . Chimeric scFv/γreceptor-mediated T cell lysis of tumor cells is co-regulated by adhesion and accessory molecules Int J Cancer 1998 77: 181–187

    Article  CAS  PubMed  Google Scholar 

  26. Willemsen RA . Grafting primary human T lymphocytes with cancer specific chimeric single chain and two chain TCR Gene Therapy 2000 7: 1369–1377

    Article  CAS  PubMed  Google Scholar 

  27. Kieke MC . Selection of functional T cell receptor mutants from a yeast surface-display library Proc Natl Acad Sci USA 1999 96: 5651–5656

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Holler PD . In vitro evolution of a T cell receptor with high affinity for peptide/MHC Proc Natl Acad Sci USA 2000 10: 5387–5392

    Article  Google Scholar 

  29. Weidanz JA . Display of functional alpha beta single-chain T cell receptor molecules on the surface of bacteriophage J Immunol Meth 1998 221: 59–76

    Article  CAS  Google Scholar 

  30. Wulfing C, Pluckthun A . Correctly folded T-cell receptor fragments in the periplasm of Escherischia coli. Influence of folding catalysis J Mol Biol 1994 5: 655–669

    Article  Google Scholar 

  31. O'Herrin SM . Analysis of the expression of peptide-major histocompatibility complexes using high affinity soluble divalent T cell receptors J Exp Med 1997 8: 1333–1345

    Article  Google Scholar 

  32. Day PM . Direct delivery of exogenous MHC class I molecule-binding oligopeptides to the endoplasmic reticulum of viable cells Proc Natl Acad Sci USA 1997 15: 8064–8069

    Article  Google Scholar 

  33. Zhong G, Reis E, Sousa C, Germain RN . Production, specificity, and functionality of monoclonal antibodies to specific peptide-major histocompatibility complex class II complexes formed by processing of exogenous protein Proc Natl Acad Sci USA 1997 94: 13856–13861

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Reiter Y . Peptide-specific killing of antigen-presenting cells by a recombinant antibody-toxin fusion protein targeted to major histocompatibility complex/peptide class I complexes with T cell receptor-like specificity Proc Natl Acad Sci USA 1997 9: 4631–4636

    Article  Google Scholar 

  35. Tamminen WL, Wraith D, Barber BH . Searching for MHC-restricted anti-viral antibodies: antibodies recognizing the nucleoprotein of influenza virus dominate the serological response of C57BL/6 mice to syngeneic influenza-infected cells Eur J Immunol 1987 7: 999–1006

    Article  Google Scholar 

  36. Chames P . Direct selection of a human antibody fragment directed against the tumor T-cell epitope HLA-A1/MAGE-A1 from a non-immunized phage-Fab library Proc Natl Acad Sci USA 2000 97: 7969–7974

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Kolb HJ, Holler E . Adoptive immunotherapy with donor lymphocyte transfusions Curr Opin Oncol 1997 9: 139–145

    Article  CAS  PubMed  Google Scholar 

  38. Garboczi DN, Hung DT, Wiley DC . HLA-A2-peptide complexes: refolding and crystallization of molecules expressed in Escherichia coli and complexed with single antigenic peptides Proc Natl Acad Sci USA 1992 89: 3429–3433

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. DuBridge RB . Analysis of mutation in human cells by using an Epstein–Barr virus shuttle system Mol Cell Biol 1987 7: 379–387

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Van de Griend RJ . Rapid expansion of human cytotoxic T cell clones: growth promotion by a heat-labile serum component and by various types of feeder cells J Immunol Meth 1984 66: 285–298

    Article  CAS  Google Scholar 

  41. Van de Griend RJ, Bolhuis RLH . Rapid expansion of allospecific cytotoxic T cell clones using nonspecific feeder cell lines without further addition of exogenous IL2 Transplantation 1984 38: 401–406

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by EU grant BIO4-CT97-2196 to PC and HRH, EU grant No. QLK3-1999-01262 and a grant from the Dutch Technology Foundation STW (project RGN44.3498) to RLHB.

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Authors and Affiliations

  1. Department of Clinical and Tumor Immunology, Academic Hospital Rotterdam/Daniel den Hoed Cancer Center, Rotterdam, The Netherlands

    RA Willemsen, R Debets, E Hart & RLH Bolhuis

  2. Department of Pathology, Maastricht University, Maastricht, The Netherlands

    HR Hoogenboom & P Chames

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  1. RA Willemsen
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  2. R Debets
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  3. E Hart
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  4. HR Hoogenboom
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Willemsen, R., Debets, R., Hart, E. et al. A phage display selected Fab fragment with MHC class I-restricted specificity for MAGE-A1 allows for retargeting of primary human T lymphocytes. Gene Ther 8, 1601–1608 (2001). https://doi.org/10.1038/sj.gt.3301570

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