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

Dolton, G., Lissina, A., Skowera, A., Ladell, K., Tungatt, K., Jones, E., Kronenberg-Versteeg, D., Akpovwa, H., Pentier, J.M., Holland, C.J., Godkin, A.J., Cole, D.K., Neller, M.A., Miles, J.J., Price, D.A., Peakman, M., and Sewell, A.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.

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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 ID: 45059
Item Type: Article (Research - C1)
ISSN: 1365-2249
Keywords: autoimmunity, diabetes, T cell receptors, T cells, tumour immunology
Additional Information:

© 2014 The Authors. Clinical & Experimental Immunology published by John Wiley & Sons Ltd on behalf of British Society for Immunology, Clinical and Experimental Immunology, 177:47–63.

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

https://creativecommons.org/licenses/by/3.0/

Funders: Juvenile Diabetes Research Foundation (JDRF), UK Department of Health (UKDH), National Institute for Health Research (NIHR), King's College, London (KCL), Australian National Health and Medical Research Council (NHMRC), Wellcome Trust (WT)
Projects and Grants: JDRF award 17-2012-352
Date Deposited: 09 Aug 2016 05:17
FoR Codes: 11 MEDICAL AND HEALTH SCIENCES > 1107 Immunology > 110702 Applied Immunology (incl Antibody Engineering, Xenotransplantation and T-cell Therapies) @ 100%
SEO Codes: 92 HEALTH > 9201 Clinical Health (Organs, Diseases and Abnormal Conditions) > 920108 Immune System and Allergy @ 100%
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