More tricks with tetramers: a practical guide to staining T cells with peptide-MHC multimers

Dolton, Garry, Tungatt, Katie, Lloyd, Angharad, Bianchi, Valentina, Theaker, Sarah M., Trimby, Andrew, Holland, Christopher J., Donia, Marco, Godkin, Andrew J., Cole, David K., Straten, Per Thor, Peakman, Mark, Svane, Marie, and Sewell, Andrew K. (2015) More tricks with tetramers: a practical guide to staining T cells with peptide-MHC multimers. Immunology, 146. pp. 11-22.

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Abstract

Analysis of antigen-specific T-cell populations by flow cytometry with peptide-MHC (pMHC) multimers is now commonplace. These reagents allow the tracking and phenotyping of T cells during infection, autoimmunity and cancer, and can be particularly revealing when used for monitoring therapeutic interventions. In 2009, we reviewed a number of 'tricks' that could be used to improve this powerful technology. More recent advances have demonstrated the potential benefits of using higher order multimers and of 'boosting' staining by inclusion of an antibody against the pMHC multimer. These developments now allow staining of T cells where the interaction between the pMHC and the T-cell receptor is over 20-fold weaker (K(D) > 1 mm) than could previously be achieved. Such improvements are particularly relevant when using pMHC multimers to stain anti-cancer or autoimmune T-cell populations, which tend to bear lower affinity T-cell receptors. Here, we update our previous work to include discussion of newer tricks that can produce substantially brighter staining even when using log-fold lower concentrations of pMHC multimer. We further provide a practical guide to using pMHC multimers that includes a description of several common pitfalls and how to circumvent them.

Item ID: 55335
Item Type: Article (Research - C1)
ISSN: 1365-2567
Funders: Juvenile Diabetes Research Foundation (JDRF), Cardiff University (CU), Wellcome Trust
Projects and Grants: JDRF 17-2012-352
Date Deposited: 23 Oct 2018 03:35
FoR Codes: 11 MEDICAL AND HEALTH SCIENCES > 1107 Immunology > 110704 Cellular Immunology @ 80%
11 MEDICAL AND HEALTH SCIENCES > 1107 Immunology > 110703 Autoimmunity @ 20%
SEO Codes: 97 EXPANDING KNOWLEDGE > 970111 Expanding Knowledge in the Medical and Health Sciences @ 100%
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