title: Combinatorial HLA-peptide bead libraries for high throughput identification of CD8⁺ T cell specificity.
abstract: Comprehensive antigenic characterization of a T cell population of unknown specificity is challenging. Existing MHC class I expression systems are limited by the practical difficulty of probing cell populations with an MHC class I peptide library and the cross-reactivity of T cells that are able to recognise many variants of an index peptide. Using emulsion PCR and emulsion in vitro transcription/translation of a random library of peptides conjugated to CD8-null HLA-A*0201 on beads, we probed HLA-A*0201-restricted T cells with specificity for influenza, CMV and EBV. We observed significant enrichment for sequences containing HLA-A2 anchors and correct viral fragments for all T cell populations. HLA bead display provides a novel approach to identify the specificity of T cells.
authors.affiliation: MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, NIHR Biomedical Research Centre, University of Oxford, UK.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Pan
authors.affiliation: MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, NIHR Biomedical Research Centre, University of Oxford, UK.
authors.forename: Li-Chieh
authors.initials: LC
authors.lastname: Huang
authors.affiliation: MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, NIHR Biomedical Research Centre, University of Oxford, UK.
authors.forename: Tao
authors.initials: T
authors.lastname: Dong
authors.affiliation: MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, NIHR Biomedical Research Centre, University of Oxford, UK.
authors.forename: Yanchun
authors.initials: Y
authors.lastname: Peng
authors.affiliation: MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, NIHR Biomedical Research Centre, University of Oxford, UK.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Cerundolo
authors.affiliation: MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, NIHR Biomedical Research Centre, University of Oxford, UK.
authors.forename: Simon
authors.initials: S
authors.lastname: McGowan
authors.affiliation: MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, NIHR Biomedical Research Centre, University of Oxford, UK.
authors.forename: Graham
authors.initials: G
authors.lastname: Ogg
mesh_terms: D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000821:virology; D020650:Combinatorial Chemistry Techniques; D003587:Cytomegalovirus / Q000276:immunology; D006680:HLA Antigens / Q000235:genetics / Q000276:immunology*; D004854:Herpesvirus 4, Human / Q000276:immunology; D057166:High-Throughput Screening Assays; D006801:Humans; D009975:Orthomyxoviridae / Q000276:immunology; D010446:Peptide Fragments / Q000276:immunology; D019151:Peptide Library; D010750:Phosphoproteins / Q000276:immunology; D016133:Polymerase Chain Reaction; D015534:Trans-Activators / Q000276:immunology; D014763:Viral Matrix Proteins / Q000276:immunology
chemical_list: D000956:Antigens, Viral; D006680:HLA Antigens; D010446:Peptide Fragments; D019151:Peptide Library; D010750:Phosphoproteins; C090236:SM protein, Human herpesvirus 4; D015534:Trans-Activators; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa; C448634:influenza matrix peptide (58-66)
keywords: Bead display; HLA library; T cells