title: Combinatorial tetramer staining and mass cytometry analysis facilitate T-cell epitope mapping and characterization.
abstract: It is currently not possible to predict which epitopes will be recognized by T cells in different individuals. This is a barrier to the thorough analysis and understanding of T-cell responses after vaccination or infection. Here, by combining mass cytometry with combinatorial peptide-MHC tetramer staining, we have developed a method allowing the rapid and simultaneous identification and characterization of T cells specific for many epitopes. We use this to screen up to 109 different peptide-MHC tetramers in a single human blood sample, while still retaining at least 23 labels to analyze other markers of T-cell phenotype and function. Among 77 candidate rotavirus epitopes, we identified six T-cell epitopes restricted to human leukocyte antigen (HLA)-A*0201 in the blood of healthy individuals. T cells specific for epitopes in the rotavirus VP3 protein displayed a distinct phenotype and were present at high frequencies in intestinal epithelium. This approach should be useful for the comprehensive analysis of T-cell responses to infectious diseases or vaccines.
authors.affiliation: Agency for Science, Technology and Research, Singapore Immunology Network, Singapore. evan_newell@immunol.a-star.edu.sg
authors.forename: Evan W
authors.initials: EW
authors.lastname: Newell
authors.forename: Natalia
authors.initials: N
authors.lastname: Sigal
authors.forename: Nitya
authors.initials: N
authors.lastname: Nair
authors.forename: Brian A
authors.initials: BA
authors.lastname: Kidd
authors.forename: Harry B
authors.initials: HB
authors.lastname: Greenberg
authors.forename: Mark M
authors.initials: MM
authors.lastname: Davis
mesh_terms: D000956:Antigens, Viral / Q000276:immunology; D018604:Epitope Mapping; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D005434:Flow Cytometry; D015234:HLA-A Antigens / Q000276:immunology*; D015395:Histocompatibility Antigens Class I / Q000276:immunology; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D006801:Humans; D013058:Mass Spectrometry; D010455:Peptides / Q000737:chemistry / Q000276:immunology*; D012401:Rotavirus / Q000276:immunology / Q000378:metabolism; D013601:T-Lymphocytes
chemical_list: D000956:Antigens, Viral; D018984:Epitopes, T-Lymphocyte; D015234:HLA-A Antigens; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D010455:Peptides