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.

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