Abstract
The goal of adoptive T cell therapy in cancer is to provide effective antitumor immunity by transfer of selected populations of tumor Ag-specific T cells. Transfer of T cells with high TCR avidity is critical for in vivo efficacy. In this study, we demonstrate that fluorescent peptide/MHC class I multimeric complexes incorporating mutations in the alpha3 domain (D227K/T228A) that abrogate binding to the CD8 coreceptor can be used to selectively isolate tumor Ag-specific T cells of high functional avidity from both in vitro expanded and ex vivo T cell populations. Sorting, cloning, and expansion of alpha3 domain mutant multimer-positive CD8 T cells enabled rapid selection of high avidity tumor-reactive T cell clones. Our results are relevant for ex vivo identification and isolation of T cells with potent antitumor activity for adoptive T cell therapy.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Antigens, Neoplasm / immunology
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Antigens, Neoplasm / metabolism*
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CD8-Positive T-Lymphocytes / immunology*
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CD8-Positive T-Lymphocytes / metabolism*
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Cell Adhesion / genetics
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Cell Adhesion / immunology
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Cell Line
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Cell Separation / methods
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Clone Cells
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Cytotoxicity Tests, Immunologic / methods
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Epitopes, T-Lymphocyte / genetics
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Epitopes, T-Lymphocyte / metabolism
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HLA-A2 Antigen / genetics*
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HLA-A2 Antigen / metabolism
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Humans
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MART-1 Antigen
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Melanoma / genetics
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Melanoma / immunology*
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Mutagenesis, Site-Directed*
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Neoplasm Proteins / immunology
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Neoplasm Proteins / metabolism
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Peptide Fragments / chemical synthesis
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Peptide Fragments / genetics*
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Peptide Fragments / metabolism
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Protein Binding / genetics
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Protein Binding / immunology
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Protein Structure, Tertiary / genetics
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Staining and Labeling
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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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MART-1 Antigen
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MLANA protein, human
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Neoplasm Proteins
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Peptide Fragments