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

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Wooldridge, Linda, Lissina, Anna, Cole, David K., Berg, Hugo van den, 1968-, Price, David A. and Sewell, Andrew K.. (2009) Tricks with tetramers : how to get the most from multimeric peptide-MHC. Immunology, Vol.126 (No.2). pp. 147-164. ISSN 0019-2805

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Official URL: http://dx.doi.org/10.1111/j.1365-2567.2008.02848.x

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'.

Item Type: Journal Article
Subjects: Q Science > QR Microbiology > QR180 Immunology
Divisions: Faculty of Science > Mathematics
Faculty of Science > Centre for Systems Biology
Journal or Publication Title: Immunology
Publisher: Wiley-Blackwell Publishing
ISSN: 0019-2805
Date: February 2009
Volume: Vol.126
Number: No.2
Number of Pages: 18
Page Range: pp. 147-164
Identification Number: 10.1111/j.1365-2567.2008.02848.x
Status: Peer Reviewed
Publication Status: Published
Access rights to Published version: Restricted or Subscription Access
URI: http://wrap.warwick.ac.uk/id/eprint/28768

Data sourced from Thomson Reuters' Web of Knowledge

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