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

Theaker, Sarah M., Rius Rafael, Cristina, Greenshields-Watson, Alexander, Lloyd, Angharad, Trimby, Andrew R., Fuller, Anna, Miles, John James, Cole, David, Peakman, Mark, Sewell, Andrew K. 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: 23 May 2019 05:02
URI: http://orca-mwe.cf.ac.uk/id/eprint/87394

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