Comparison of peptide-major histocompatibility complex tetramers and dextramers for the identification of antigen-specific T cells

Dolton, G., Lissina, A., Skowera, A., Ladell, K., Tungatt, K., Jones, E., Kronenberg-Versteeg, D., Akpovwa, H., Pentier, J. M., Holland, C. J., Godkin, A. J., Cole, D. K., Neller, M. A., Miles, J. J., Price, D. A., Peakman, M. and Sewell, A. K. (2014) Comparison of peptide-major histocompatibility complex tetramers and dextramers for the identification of antigen-specific T cells. Clinical and Experimental Immunology, 177 1: 47-63. doi:10.1111/cei.12339


Author Dolton, G.
Lissina, A.
Skowera, A.
Ladell, K.
Tungatt, K.
Jones, E.
Kronenberg-Versteeg, D.
Akpovwa, H.
Pentier, J. M.
Holland, C. J.
Godkin, A. J.
Cole, D. K.
Neller, M. A.
Miles, J. J.
Price, D. A.
Peakman, M.
Sewell, A. K.
Title Comparison of peptide-major histocompatibility complex tetramers and dextramers for the identification of antigen-specific T cells
Journal name Clinical and Experimental Immunology   Check publisher's open access policy
ISSN 1365-2249
0009-9104
Publication date 2014-07-01
Year available 2014
Sub-type Article (original research)
DOI 10.1111/cei.12339
Open Access Status DOI
Volume 177
Issue 1
Start page 47
End page 63
Total pages 17
Place of publication Chichester, West Sussex, United Kingdom
Publisher Wiley-Blackwell Publishing
Language eng
Abstract Fluorochrome-conjugated peptide-major histocompatibility complex (pMHC) multimers are widely used for flow cytometric visualization of antigen-specific T cells. The most common multimers, streptavidin-biotin-based 'tetramers', can be manufactured readily in the laboratory. Unfortunately, there are large differences between the threshold of T cell receptor (TCR) affinity required to capture pMHC tetramers from solution and that which is required for T cell activation. This disparity means that tetramers sometimes fail to stain antigen-specific T cells within a sample, an issue that is particularly problematic when staining tumour-specific, autoimmune or MHC class II-restricted T cells, which often display TCRs of low affinity for pMHC. Here, we compared optimized staining with tetramers and dextramers (dextran-based multimers), with the latter carrying greater numbers of both pMHC and fluorochrome per molecule. Most notably, we find that: (i) dextramers stain more brightly than tetramers; (ii) dextramers outperform tetramers when TCR-pMHC affinity is low; (iii) dextramers outperform tetramers with pMHC class II reagents where there is an absence of co-receptor stabilization; and (iv) dextramer sensitivity is enhanced further by specific protein kinase inhibition. Dextramers are compatible with current state-of-the-art flow cytometry platforms and will probably find particular utility in the fields of autoimmunity and cancer immunology.
Keyword Autoimmunity
Diabetes
T cell receptors
T cells
Tumour immunology
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: School of Medicine Publications
Official 2015 Collection
 
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