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 ORCID: https://orcid.org/0000-0002-9856-2938, Tungatt, Katie, Jones, Emma, Kronenberg-Versteeg, D., Akpovwa, Hephzibah, Pentier, Johanne, Holland, C. J., Godkin, Andrew James ORCID: https://orcid.org/0000-0002-1910-7567, Cole, David ORCID: https://orcid.org/0000-0003-0028-9396, Neller, M. A., Miles, John James, Price, David ORCID: https://orcid.org/0000-0001-9416-2737, Peakman, M. and Sewell, Andrew K. ORCID: https://orcid.org/0000-0003-3194-3135 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:	15 May 2024 01:14
URI:	https://orca.cardiff.ac.uk/id/eprint/60456

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