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

Dolton, Garry Michael, Tungatt, Katie, Lloyd, Angharad, Bianchi, Valentina, Theaker, Sarah M., Trimby, Andrew R., Holland, Christopher J., Donia, Marco, Godkin, Andrew James ORCID: https://orcid.org/0000-0002-1910-7567, Cole, David ORCID: https://orcid.org/0000-0003-0028-9396, Straten, Per Thor, Peakman, Mark, Svane, Inge Marie and Sewell, Andrew K. ORCID: https://orcid.org/0000-0003-3194-3135 2015. More tricks with tetramers: a practical guide to staining T cells with peptide-MHC multimers. Immunology 146 (1) , pp. 11-22. 10.1111/imm.12499

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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 Ab against the pMHC multimer. These developments now allow staining of T cells where the interaction between the pMHC and the T cell receptor (TCR) is over 20-fold weaker (KD > 1 mM) than could previously be achieved. Such improvements are particularly relevant when using pMHC multimers to stain anticancer or autoimmune T cell populations, which tend to bear lower affinity TCRs. 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 Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine Systems Immunity Research Institute (SIURI)
Subjects:	Q Science > QH Natural history > QH426 Genetics Q Science > QR Microbiology > QR180 Immunology
Uncontrolled Keywords:	T-cell; TCR ;peptide-MHC tetramer;peptide-MHC dextramer
Publisher:	Wiley Blackwell
ISSN:	0019-2805
Date of Acceptance:	27 May 2015
Last Modified:	15 May 2024 01:16
URI:	https://orca.cardiff.ac.uk/id/eprint/74242

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