title: T-cell libraries allow simple parallel generation of multiple peptide-specific human T-cell clones.
abstract: Isolation of peptide-specific T-cell clones is highly desirable for determining the role of T-cells in human disease, as well as for the development of therapies and diagnostics. However, generation of monoclonal T-cells with the required specificity is challenging and time-consuming. Here we describe a library-based strategy for the simple parallel detection and isolation of multiple peptide-specific human T-cell clones from CD8(+) or CD4(+) polyclonal T-cell populations. T-cells were first amplified by CD3/CD28 microbeads in a 96U-well library format, prior to screening for desired peptide recognition. T-cells from peptide-reactive wells were then subjected to cytokine-mediated enrichment followed by single-cell cloning, with the entire process from sample to validated clone taking as little as 6 weeks. Overall, T-cell libraries represent an efficient and relatively rapid tool for the generation of peptide-specific T-cell clones, with applications shown here in infectious disease (Epstein-Barr virus, influenza A, and Ebola virus), autoimmunity (type 1 diabetes) and cancer.
authors.affiliation: Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, UK.
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Theaker
authors.affiliation: Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, UK.
authors.forename: Cristina
authors.initials: C
authors.lastname: Rius
authors.affiliation: Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, UK.
authors.forename: Alexander
authors.initials: A
authors.lastname: Greenshields-Watson
authors.affiliation: Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, UK.
authors.forename: Angharad
authors.initials: A
authors.lastname: Lloyd
authors.affiliation: Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, UK.
authors.forename: Andrew
authors.initials: A
authors.lastname: Trimby
authors.affiliation: Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, UK.
authors.forename: Anna
authors.initials: A
authors.lastname: Fuller
authors.affiliation: Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, UK; QIMR Berghofer Medical Research Institute, Brisbane, Queensland 4029, Australia.
authors.forename: John J
authors.initials: JJ
authors.lastname: Miles
authors.affiliation: Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, UK.
authors.forename: David K
authors.initials: DK
authors.lastname: Cole
authors.affiliation: Peter Gorer Department of Immunobiology, King's College London Faculty of Life Sciences & Medicine, Guy's Hospital, London, UK.
authors.forename: Mark
authors.initials: M
authors.lastname: Peakman
authors.affiliation: Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, UK. Electronic address: sewellak@cf.ac.uk.
authors.forename: Andrew K
authors.initials: AK
authors.lastname: Sewell
authors.affiliation: Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, UK.
authors.forename: Garry
authors.initials: G
authors.lastname: Dolton
mesh_terms: D000956:Antigens, Viral / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002999:Clone Cells / Q000276:immunology; D003602:Cytotoxicity, Immunologic; D029043:Ebolavirus / Q000276:immunology; D058501:Enzyme-Linked Immunospot Assay / Q000379:methods; D004854:Herpesvirus 4, Human; D006801:Humans; D010455:Peptides / Q000235:genetics* / Q000276:immunology*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology
chemical_list: D000956:Antigens, Viral; D010455:Peptides; D016693:Receptors, Antigen, T-Cell, alpha-beta
keywords: Ebola; Library; Peptide-specific; T-cell clone; Tumour; Type 1 diabetes
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