Journal
IMMUNOLOGY
Volume 146, Issue 1, Pages 11-22Publisher
WILEY
DOI: 10.1111/imm.12499
Keywords
peptide-MHC dextramer; peptide-MHC tetramer; T-cell receptor; T cell
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Funding
- Juvenile Diabetes Research Foundation [JDRF 17-2012-352]
- Cardiff University
- Biotechnology and Biological Sciences grant [BB/H001085/1]
- MRC
- Cancer Research Wales studentship
- Breast Cancer Now
- BBSRC [BB/H001085/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/H001085/1] Funding Source: researchfish
- Medical Research Council [1247762] Funding Source: researchfish
- The Danish Cancer Society [R72-A4396] Funding Source: researchfish
- Wellcome Trust [100327/Z/12/Z] Funding Source: researchfish
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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>1mm) 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.
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