Tetramer-guided analysis of TCR beta-chain usage reveals a large repertoire of melan-A-specific CD8+ T cells in melanoma patients.

Abstract

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.

• Authors: D Valmori, V Dutoit, D Liénard, F Lejeune, D Speiser, D Rimoldi, V Cerundolo, P Y Dietrich, J C Cerottini, P Romero
• Journal: Journal of immunology (Baltimore, Md. : 1950) (J Immunol) Vol. 165 Issue 1 Pg. 533-8 (Jul 01 2000) ISSN: 0022-1767 [Print] United States
• PMID: 10861093 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antigens, Neoplasm
• Epitopes, T-Lymphocyte
• MART-1 Antigen
• MLANA protein, human
• Neoplasm Proteins
• Peptide Fragments

Topics

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