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Previous attempts to treat human malignancies by adoptive transfer of tumor-specific CTLs have been limited by the difficulty of isolating T cells of defined antigen specificity. The recent development of MHC class I/antigenic peptide tetrameric complexes that allow direct identification of antigen-specific T cells has opened new possibilities for the isolation and in vitro expansion of tumor-specific T cells. In the present study, we have derived polyclonal monospecific cell lines from circulating Melan-A-specific CTL precursors of HLA-A*0201+ melanoma patients by combining stimulation with recently identified peptide analogues of the immunodominant epitope from the melanoma-associated antigen Melan-A with staining with fluorescent HLA-A*0201/Melan-A peptide tetramers. In vitro expansion of antigen-specific CD8+ T cells was monitored by flow cytometry with the fluorescent tetramers and anti-CD8 monoclonal antibody. This analysis revealed that Melan-A 26-35 peptide analogues were much more efficient than the parental peptides in stimulating a rapid in vitro expansion of antigen-specific CD8+ T cells. These cells were then isolated by tetramer-guided cell sorting and subsequently expanded in vitro by mitogen stimulation. The resulting polyclonal but monospecific CTLs fully cross-recognized the parental peptides and were able to efficiently lyse Melan-A-expressing tumor cells. Altogether, these results pave the way to a molecularly defined approach to antigen-specific adoptive transfer therapy of cancer.


Journal article


Cancer research

Publication Date





2167 - 2173


Division of Clinical Onco-Immunology, Ludwig Institute for Cancer Research, University Hospital, CHUV, Lausanne, Switzerland.


T-Lymphocytes, Cytotoxic, Cell Line, Tumor Cells, Cultured, Humans, Melanoma, Macromolecular Substances, Peptide Fragments, Neoplasm Proteins, Recombinant Fusion Proteins, Interleukin-2, Antigens, Neoplasm, HLA-A2 Antigen, Immunodominant Epitopes, Fluorescent Antibody Technique, Indirect, Immunotherapy, Adoptive, Flow Cytometry, Cell Separation, Biotinylation, Lymphocyte Activation, Cytotoxicity, Immunologic, MART-1 Antigen