T-cell libraries allow simple parallel generation of multiple peptide-specific human T-cell clones

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:	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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