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Comparison of peptide-major histocompatibility complex tetramers and dextramers for the identification of antigen-specific T cells

Dolton, Garry Michael, Lissina, A., Skowera, A., Ladell, Kristin Ingrid, Tungatt, Katie, Jones, Emma, Kronenberg-Versteeg, D., Akpovwa, Hephzibah, Pentier, Johanne, Holland, C. J., Godkin, Andrew James, Cole, David, Neller, M. A., Miles, John James, Price, David, Peakman, M. and Sewell, Andrew K. 2014. Comparison of peptide-major histocompatibility complex tetramers and dextramers for the identification of antigen-specific T cells. Clinical and Experimental Immunology 177 (1) , pp. 47-63. 10.1111/cei.12339

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Abstract

Fluorochrome-conjugated peptide–major histocompatibility complex (pMHC) multimers are widely used for flow cytometric visualization of antigen-specific T cells. The most common multimers, streptavidin–biotin-based ‘tetramers’, can be manufactured readily in the laboratory. Unfortunately, there are large differences between the threshold of T cell receptor (TCR) affinity required to capture pMHC tetramers from solution and that which is required for T cell activation. This disparity means that tetramers sometimes fail to stain antigen-specific T cells within a sample, an issue that is particularly problematic when staining tumour-specific, autoimmune or MHC class II-restricted T cells, which often display TCRs of low affinity for pMHC. Here, we compared optimized staining with tetramers and dextramers (dextran-based multimers), with the latter carrying greater numbers of both pMHC and fluorochrome per molecule. Most notably, we find that: (i) dextramers stain more brightly than tetramers; (ii) dextramers outperform tetramers when TCR–pMHC affinity is low; (iii) dextramers outperform tetramers with pMHC class II reagents where there is an absence of co-receptor stabilization; and (iv) dextramer sensitivity is enhanced further by specific protein kinase inhibition. Dextramers are compatible with current state-of-the-art flow cytometry platforms and will probably find particular utility in the fields of autoimmunity and cancer immunology.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
Systems Immunity Research Institute (SIURI)
Subjects: R Medicine > R Medicine (General)
Publisher: Wiley-Blackwell
ISSN: 0009-9104
Funders: Wellcome Trust
Date of First Compliant Deposit: 30 March 2016
Date of Acceptance: 12 March 2014
Last Modified: 28 Jun 2019 02:03
URI: http://orca-mwe.cf.ac.uk/id/eprint/60456

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