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In situ detection of antigen-specific T cells in cryo-sections using MHC class I tetramers after dendritic cell vaccination of melanoma patients

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Abstract

Application of tetrameric MHC class I-peptide complexes has significantly improved the monitoring of antigen-specific T cell immune responses in mouse models as well as in clinical studies. Especially MHC class I tetramer analysis of tumor-specific T cells in suspension or on thick vibratome sections from viable tissue has been proven extremely useful. Using the well-characterized mouse tyrosinase-related-protein-2 specific cytotoxic T cell (CTL) clone LP9, we now developed a method that allows for specific identification of T cells with MHC class I tetramers in 8 μm thick, chemically fixed cryosections. The protocol was validated in a murine influenza virus-infection model. Moreover, analysis of delayed type hypersensitivity (DTH) skin biopsies from melanoma patients vaccinated with peptide-loaded mature dendritic cells, revealed the presence and location of anti-tumor CTLs. The specificity of the CTLs detected in situ correlated with both the DTH challenge specificity and reactivity of cell suspensions derived from the same biopsies. Collectively, our data demonstrate that in situ MHC class I tetramer staining provides a valuable tool to reveal the presence and anatomical location of specific CTLs in frozen tissue following immune-based treatment strategies in cancer patients.

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Acknowledgments

We are grateful to F. A. Vyth-Dreese, K. Schepers H. Spits and T. Schumacher for providing precious material and for helpful suggestions. This study was supported by grants KUN 1995/1910, 1999/1950, 2000/2301, 2003/2893 and 2003/2917 from the Dutch Cancer Society and the TIL-foundation.

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Authors and Affiliations

  1. Department of Tumor Immunology, Radboud University Nijmegen Medical Centre, Nijmegen Centre for Molecular Life Sciences, Nijmegen, The Netherlands

    N. M. Scharenborg, C. G. Figdor & G. J. Adema

  2. Department of Tumor Immunology, NCMLS/278 TIL, University Medical Centre, Postbox 9101, 6500 HB Nijmegen, The Netherlands

    G. J. Adema

  3. Department of Pathology, Radboud University Nijmegen Medical Centre, Nijmegen Centre for Molecular Life Sciences, Nijmegen, The Netherlands

    M. R. Bernsen, W. L. van Geloof, P. D. M. Rombout, G. N. P. Van Muijen & D. J. Ruiter

  4. Department of Tumor Immunology, Medical Oncology and Pediatric Hemato-Oncology, Radboud University Nijmegen Medical Centre, Nijmegen Centre for Molecular Life Sciences, Nijmegen, The Netherlands

    I. J. M. De Vries

  5. Department of Medical Oncology, Radboud University Nijmegen Medical Centre, Nijmegen Centre for Molecular Life Sciences, Nijmegen, The Netherlands

    W. J. Lesterhuis & C. J. A. Punt

Authors
  1. I. J. M. De Vries
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  2. M. R. Bernsen
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  3. W. L. van Geloof
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  4. N. M. Scharenborg
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  5. W. J. Lesterhuis
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  6. P. D. M. Rombout
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  7. G. N. P. Van Muijen
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  8. C. G. Figdor
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  9. C. J. A. Punt
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  10. D. J. Ruiter
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  11. G. J. Adema
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Corresponding author

Correspondence to G. J. Adema.

Additional information

De Vries and Bernsen contributed equally to this study.

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De Vries, I.J.M., Bernsen, M.R., van Geloof, W.L. et al. In situ detection of antigen-specific T cells in cryo-sections using MHC class I tetramers after dendritic cell vaccination of melanoma patients. Cancer Immunol Immunother 56, 1667–1676 (2007). https://doi.org/10.1007/s00262-007-0304-5

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  • DOI: https://doi.org/10.1007/s00262-007-0304-5

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