Abstract
Ex vivo ELISPOT analysis of peripheral blood lymphocytes obtained from stage IV melanoma patients demonstrated reactivity against peptides derived from MART-1 and gp100. However, the number of reactive T cells was < 1% that of total lymphocytes as detected by flow cytometry using tetrameric MHC/peptide complexes. Despite this low frequency, we were able to directly isolate these populations ex vivo by means of magnetic beads coated with MHC/peptide complexes and to subject these cells to T-cell receptor clonotype mapping. This analysis revealed that the MART-1/A*0201- and gp100/A*0201-reactive T-cell populations are composed of oligoclonal T cells that engage several T-cell receptor beta chain families. Longitudinal studies using this approach may result in a better correlation between T-cell reactivity and the course of neoplastic disease.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Aged
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Antigens, Neoplasm
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Biomarkers, Tumor / immunology
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Clone Cells
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Electrophoresis, Polyacrylamide Gel
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Epitopes, T-Lymphocyte / immunology
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Flow Cytometry
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HLA-A Antigens / immunology
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Humans
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MART-1 Antigen
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Male
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Melanoma / immunology*
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Melanoma / pathology
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Membrane Glycoproteins / immunology
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Middle Aged
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Neoplasm Proteins / immunology*
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Neoplasm Staging
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RNA, Neoplasm / genetics
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RNA, Neoplasm / metabolism
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Receptors, Antigen, T-Cell / genetics
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Receptors, Antigen, T-Cell / immunology*
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Receptors, Antigen, T-Cell, alpha-beta / genetics
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Receptors, Antigen, T-Cell, alpha-beta / immunology
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Reverse Transcriptase Polymerase Chain Reaction
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T-Lymphocytes / cytology
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T-Lymphocytes / immunology*
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gp100 Melanoma Antigen
Substances
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Antigens, Neoplasm
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Biomarkers, Tumor
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Epitopes, T-Lymphocyte
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HLA-A Antigens
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MART-1 Antigen
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MLANA protein, human
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Membrane Glycoproteins
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Neoplasm Proteins
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PMEL protein, human
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RNA, Neoplasm
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Receptors, Antigen, T-Cell
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Receptors, Antigen, T-Cell, alpha-beta
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gp100 Melanoma Antigen