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Analysis of T-cell responses in metastatic melanoma patients vaccinated with dendritic cells pulsed with tumor lysates

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Abstract

In melanoma patients, CD8+ cytotoxic T cells have been found recognizing self-proteins of which the expression is restricted to the melanocytic lineage. These melanocyte differentiation antigens are expressed in normal melanocytes as well as in 80–100% of primary and metastatic melanoma. In this report, six HLA-A*0201–subtyped metastatic melanoma patients vaccinated with dendritic cells (DCs) pulsed with autologous tumor lysates and keyhole limpet hemocyanin (KLH) were screened for the presence of CD8+ T cells specific for three HLA-A*0201–binding peptides derived from the melanosomal antigens MART-1/Melan-A, gp100, and tyrosinase. For this purpose, nonstimulated as well as in vitro peptide-stimulated peripheral blood mononuclear cells (PBMCs) were tested for peptide-specific IFN-γ release by enzyme-linked immunosorbent spot (ELISpot) assays. Furthermore, expression of the melanosomal antigens MART-1/Melan-A, gp100, and tyrosinase in tumor lesions was analyzed by immunohistochemistry before and after vaccination. We also used the ELISpot technique to investigate whether KLH-specific T cells were induced and whether these cells released type 1 (IFN-γ) and/or type 2 (IL-13) cytokines. Our data show induction of CD8+ T cells specific for the melanosomal peptides MART-1/Melan-A27–35 or tyrosinase1–9, as well as IFN-γ–releasing KLH-specific T cells, in two of six vaccinated melanoma patients, but do not support an association between the induction of these T cells and clinical responses.

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Acknowledgements

This work was supported by a grant from the Dutch Cancer Society (NKB 95-1354).

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Authors and Affiliations

  1. Department of Clinical Oncology, Leiden University Medical Center, Leiden, The Netherlands

    Marieke Griffioen, Martina Borghi, Peter I. Schrier & Susanne Osanto

  2. Skin Cancer Unit, German Cancer Research Center and Department of Dermatology, University Hospital Mannheim, Universität Heidelberg, Theodor Kutzer Ufer 1, 68167, Mannheim, Germany

    Dirk Schadendorf

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  1. Marieke Griffioen
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  2. Martina Borghi
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  3. Peter I. Schrier
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  4. Susanne Osanto
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  5. Dirk Schadendorf
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Correspondence to Dirk Schadendorf.

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Griffioen, M., Borghi, M., Schrier, P.I. et al. Analysis of T-cell responses in metastatic melanoma patients vaccinated with dendritic cells pulsed with tumor lysates. Cancer Immunol Immunother 53, 715–722 (2004). https://doi.org/10.1007/s00262-004-0514-z

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  • DOI: https://doi.org/10.1007/s00262-004-0514-z

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