Theaker, Sarah M., Rius Rafael, Cristina  ORCID: https://orcid.org/0000-0002-1896-1649, Greenshields-Watson, Alexander, Lloyd, Angharad, Trimby, Andrew R., Fuller, Anna, Miles, John James, Cole, David ORCID: https://orcid.org/0000-0003-0028-9396, Peakman, Mark, Sewell, Andrew K. ORCID: https://orcid.org/0000-0003-3194-3135 and Dolton, Garry Michael
2016.
T-cell libraries allow simple parallel generation of multiple peptide-specific human T-cell clones.
Journal of Immunological Methods
430
, pp. 43-50.
10.1016/j.jim.2016.01.014
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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 Type: | Article |
|---|---|
| Date Type: | Publication |
| Status: | Published |
| Schools: | Schools > Medicine |
| Subjects: | Q Science > QR Microbiology > QR180 Immunology |
| Publisher: | Elsevier |
| ISSN: | 0022-1759 |
| Funders: | Wellcome Trust |
| Date of First Compliant Deposit: | 30 March 2016 |
| Date of Acceptance: | 26 January 2016 |
| Last Modified: | 15 May 2024 01:18 |
| URI: | https://orca.cardiff.ac.uk/id/eprint/87394 |
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