Abstract
Presentation of intracellular tumor-associated Ags (TAAs) in the context of HLA class I molecules offers unique cancer-specific cell surface markers for the identification and targeting of tumor cells. For most peptide Ags, the levels of and variations in cell surface presentation remain unknown, yet these parameters are of crucial importance when considering specific TAAs as targets for anticancer therapy. Here we use a soluble TCR with picomolar affinity for the HLA-A2-restricted 157-165 epitope of the NY-ESO-1 and LAGE-1 TAAs to investigate presentation of this immunodominant epitope on the surface of a variety of cancer cells. By single molecule fluorescence microscopy, we directly visualize HLA-peptide presentation for the first time, demonstrating that NY-ESO-1/LAGE-1-positive tumor cells present 10-50 NY-ESO-1/LAGE-1(157-165) epitopes per cell.
MeSH terms
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Amino Acid Sequence
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Antigen Presentation
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Antigen-Presenting Cells / immunology
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Antigen-Presenting Cells / metabolism
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Antigen-Presenting Cells / pathology
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Antigens, Neoplasm / analysis*
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Antigens, Neoplasm / biosynthesis
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Antigens, Neoplasm / metabolism
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Antigens, Surface
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Cell Line, Tumor
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Cytotoxicity, Immunologic
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Epitopes, T-Lymphocyte / analysis*
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Epitopes, T-Lymphocyte / biosynthesis
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Epitopes, T-Lymphocyte / metabolism
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HCT116 Cells
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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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Immunodominant Epitopes / analysis
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Immunodominant Epitopes / biosynthesis
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Immunodominant Epitopes / metabolism
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Immunosuppressive Agents / metabolism
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Melanoma / immunology
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Melanoma / metabolism
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Membrane Proteins / analysis*
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Membrane Proteins / biosynthesis
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Membrane Proteins / metabolism
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Molecular Sequence Data
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Peptide Fragments / analysis*
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Peptide Fragments / biosynthesis
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Peptide Fragments / immunology
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Peptide Fragments / metabolism
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Protein Binding
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Receptors, Antigen, T-Cell / metabolism*
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Receptors, Antigen, T-Cell / physiology
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T-Lymphocytes, Cytotoxic / immunology
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T-Lymphocytes, Cytotoxic / metabolism
Substances
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Antigens, Neoplasm
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Antigens, Surface
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CTAG1B protein, human
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CTAG2 protein, human
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Epitopes, T-Lymphocyte
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HLA-A2 Antigen
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Immunodominant Epitopes
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Immunosuppressive Agents
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Membrane Proteins
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Peptide Fragments
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Receptors, Antigen, T-Cell