Abstract
Avidity of Ag recognition by tumor-specific T cells is one of the main parameters that determines the potency of a tumor rejection Ag. In this study we show that the relative efficiency of staining of tumor Ag-specific T lymphocytes with the corresponding fluorescent MHC class I/peptide multimeric complexes can considerably vary with staining conditions and does not necessarily correlate with avidity of Ag recognition. Instead, we found a clear correlation between avidity of Ag recognition and the stability of MHC class I/peptide multimeric complexes interaction with TCR as measured in dissociation kinetic experiments. These findings are relevant for both identification and isolation of tumor-reactive CTL.
MeSH terms
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Antigens, Neoplasm / immunology
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Antigens, Neoplasm / metabolism*
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Clone Cells
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Cytotoxicity Tests, Immunologic / methods
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Cytotoxicity, Immunologic*
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Epitopes, T-Lymphocyte / immunology
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Epitopes, T-Lymphocyte / metabolism*
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Flow Cytometry / methods
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HLA-A2 Antigen / immunology
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HLA-A2 Antigen / metabolism*
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Humans
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Kinetics
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Peptide Fragments / immunology
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Peptide Fragments / metabolism*
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Protein Binding / immunology
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Receptors, Antigen, T-Cell / immunology
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Receptors, Antigen, T-Cell / metabolism*
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Staining and Labeling
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T-Lymphocytes, Cytotoxic / immunology
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T-Lymphocytes, Cytotoxic / metabolism*
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Tumor Cells, Cultured
Substances
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Antigens, Neoplasm
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Epitopes, T-Lymphocyte
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HLA-A2 Antigen
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Peptide Fragments
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Receptors, Antigen, T-Cell