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The assessment of the TCR repertoire expressed by tumor-specific CD8+ T lymphocytes has been hampered to date by the difficulty of targeting the analysis to lymphocytes directed against a single epitope. In the present study we have used fluorescent A2/Melan-A tetramers in conjunction with anti-CD8 and anti-TCR beta-chain variable (BV) mAbs to analyze by flow cytometry the BV segment usage by Melan-A-specific CD8+ T cells in tumor-infiltrated lymph nodes (TILN) and tumor-infiltrating lymphocytes (TIL) from A2 melanoma patients. Analysis of TILN populations revealed small proportions of A2/Melan-A tetramer+ cells expressing many different BV together with over-representation of A2/Melan-A tetramer+ cells expressing certain BVs. The BV usage by A2/Melan-A tetramer+ lymphocytes in TIL was more restricted than that in TILN. Moreover, the predominant BV segments were quite distinct in populations derived from different patients. A2/Melan-A tetramer+ cells expressing the dominant BVs found in TILN could also be found in the corresponding peptide-stimulated autologous PBMC, although A2/Melan-A tetramer+ lymphocytes expressing additional BVs were also identified. Together, these results suggest that a large and diverse repertoire of Melan-A-specific T cells using different BV TCR segments is available in A2 melanoma patients.


Journal article


J Immunol

Publication Date





533 - 538


Adult, Aged, Antigens, Neoplasm, CD8-Positive T-Lymphocytes, Epitopes, T-Lymphocyte, Gene Rearrangement, beta-Chain T-Cell Antigen Receptor, Genes, T-Cell Receptor beta, Humans, Leukocytes, Mononuclear, Lymphocyte Activation, Lymphocyte Count, Lymphocytes, Tumor-Infiltrating, MART-1 Antigen, Melanoma, Middle Aged, Neoplasm Proteins, Peptide Fragments, Tumor Cells, Cultured