T-cell libraries allow simple parallel generation of multiple peptide-specific human T-cell clones
Theaker, Sarah M., Rius, Cristina, Greenshields-Watson, Alexander, Lloyd, Angharad, Trimby, Andrew, Fuller, Anna, Miles, John J., Cole, David K., Peakman, Mark, Sewell, Andrew K., and Dolton, Garry (2016) T-cell libraries allow simple parallel generation of multiple peptide-specific human T-cell clones. Journal of Immunological Methods, 430. pp. 43-50.
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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.
Item ID: | 44950 |
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Item Type: | Article (Research - C1) |
ISSN: | 1872-7905 |
Keywords: | Ebola; library; peptide-specific; T-cell clone; tumour; type 1 diabetes |
Additional Information: | © 2016 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). Open Access funded by Wellcome Trust. |
Funders: | Breast Cancer Now (BCN), Cancer Research Wales (CRW), Life Sciences Research Network Wales (LSRNW), MRC, National Health and Medical Research Council (NHMRC), Wellcome Trust (WT), National Institute for Health Research Biomedical Research Centre (NIHRBRC) |
Date Deposited: | 02 Aug 2016 23:04 |
FoR Codes: | 32 BIOMEDICAL AND CLINICAL SCIENCES > 3204 Immunology > 320402 Applied immunology (incl. antibody engineering, xenotransplantation and t-cell therapies) @ 100% |
SEO Codes: | 92 HEALTH > 9201 Clinical Health (Organs, Diseases and Abnormal Conditions) > 920108 Immune System and Allergy @ 100% |
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