Parallel detection of antigen-specific T-cell responses by multidimensional encoding of MHC multimers

• Published in: Nature methods Vol. 6; no. 7; pp. 520 - 526
• Main Authors: Hadrup, Sine Reker, Bakker, Arnold H, Shu, Chengyi J, Andersen, Rikke S, van Veluw, Jerre, Hombrink, Pleun, Castermans, Emilie, thor Straten, Per, Blank, Christian, Haanen, John B, Heemskerk, Mirjam H, Schumacher, Ton N
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.07.2009; Nature Publishing Group
• Subjects: Antigen (tumor-associated); Antigen receptors, T cell; Antigenic determinants; Antigens; Antigens, Neoplasm; Assaying; Bioinformatics; Biological Microscopy; Biological Techniques; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Blood; Cell populations; Cell Separation - methods; Cellular biology; Combinatorial analysis; Epitopes; Fluorescence; Fluorescent Dyes; Health aspects; Histocompatibility antigen HLA; Histocompatibility Antigens - chemistry; Histocompatibility Antigens - metabolism; Humans; Identification and classification; Immune response; Immune system; Immunity; Immunochemistry; Immunologic Techniques; Immunological research; Leukocytes; Life Sciences; Lymphocytes; Lymphocytes T; Major histocompatibility complex; Melanoma; Melanoma-Specific Antigens; Methods; Molecular biology; Nanotechnology; Neoplasm Proteins; Peptides - immunology; Peripheral blood; Physiological aspects; Populations; Properties; Protein Structure, Quaternary; Proteomics; Quantum Dots; Receptors; Scientific method; Sensitivity and Specificity; Stains and staining (Microscopy); T cells; T-Lymphocyte Subsets - classification; T-Lymphocyte Subsets - cytology; T-Lymphocyte Subsets - immunology; Netherlands
• ISSN: 1548-7091; 1548-7105; 1548-7105
• DOI: 10.1038/nmeth.1345

Using combinations of fluorescently labeled peptide–major histocompatability complex (pMHC) tetramers, T-cell populations with multiple antigen specificities can be monitored in parallel from small samples of human blood. Also in this issue, Newell et al . present a very similar combinatorial encoding method for this purpose. The use of fluorescently labeled major histocompatibility complex multimers has become an essential technique for analyzing disease- and therapy-induced T-cell immunity. Whereas classical major histocompatibility complex multimer analyses are well-suited for the detection of immune responses to a few epitopes, limitations on human-subject sample size preclude a comprehensive analysis of T-cell immunity. To address this issue, we developed a combinatorial encoding strategy that allows the parallel detection of a multitude of different T-cell populations in a single sample. Detection of T cells from peripheral blood by combinatorial encoding is as efficient as detection with conventionally labeled multimers but results in a substantially increased sensitivity and, most notably, allows comprehensive screens to be performed. We obtained proof of principle for the feasibility of large-scale screening of human material by analysis of human leukocyte antigen A3–restricted T-cell responses to known and potential melanoma-associated antigens in peripheral blood from individuals with melanoma.