Tricks with tetramers: how to get the most from multimeric peptide-MHC

Anya Lissina, Linda Wooldridge, David K Cole, Hugo A van den Berg, David A Price, Andrew K Sewell

Research output: Contribution to journalArticle (Academic Journal)peer-review

110 Citations (Scopus)

Abstract

The development of fluorochrome-conjugated peptide-major histocompatibility complex (pMHC) multimers in conjunction with continuing advances in flow cytometry has transformed the study of antigen-specific T cells by enabling their visualization, enumeration, phenotypic characterization and isolation from ex vivo samples. Here, we bring together and discuss some of the 'tricks' that can be used to get the most out of pMHC multimers. These include: (1) simple procedures that can substantially enhance the staining intensity of cognate T cells with pMHC multimers; (2) the use of pMHC multimers to stain T cells with very-low-affinity T-cell receptor (TCR)/pMHC interactions, such as those that typically predominate in tumour-specific responses; and (3) the physical grading and clonotypic dissection of antigen-specific T cells based on the affinity of their cognate TCR using mutant pMHC multimers in conjunction with new approaches to the molecular analysis of TCR gene expression. We also examine how soluble pMHC can be used to examine T-cell activation, manipulate T-cell responses and study allogeneic and superantigen interactions with TCRs. Finally, we discuss the problems that arise with pMHC class II (pMHCII) multimers because of the low affinity of TCR/pMHCII interactions and lack of 'coreceptor help'.

Original languageEnglish
Pages (from-to)147-64
Number of pages18
JournalImmunology
Volume126
Issue number2
DOIs
Publication statusPublished - Feb 2009

Keywords

  • Autoantigens
  • Epitopes, T-Lymphocyte
  • Flow Cytometry
  • Humans
  • Lymphocyte Activation
  • Lymphocytes, Tumor-Infiltrating
  • Major Histocompatibility Complex
  • Peptides
  • Receptors, Antigen, T-Cell
  • Staining and Labeling
  • T-Lymphocyte Subsets

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