Performance of MHC Dextramer™ Reagents
MHC Dextramer™ reagents are used in the study of antigen-specific T cells. High avidity, brightness and excellent resolution make MHC Dextramer™ reagents the superior choice for detection of antigen-specific T-cells in flow cytometry and immunohistochemistry.
Flow cytometry
The MHC Dextramer™ reagents come with any of three different flurochromes
(PE, APC or FITC). When using flow cytometry they may be used to accurately
monitor CD8+ T-cell responses in blood, CSF or other fluid cell samples.
Below the benefits of using Dextramer™ reagents compared to conventional MHC multimers are shown.
MHC Dextramers can detect low affinity antigen-specific T cells, Tetramers cannot
(Preliminary data kindly provided by independent UK lab)
A, B, and C: Flow cytometry analysis of a cell sample constituting 99% “negative” HPBMC cells spiked with 1% cells of a T- cell line specific for the multimer reagents. Cells were stained with either Tetramers or Dextramers carrying pMHC with A) low affinity for TCR, B) medium affinity for TCR, or C) no affinity for TCR (negative control). Gated cells; live singlets, CD3+, CD14- and CD19-. D) Signal intensity of the T-cell line when stained with the respective MHC multimers. Cell count and multimer signal is shown for each of the three multimer specificities. Given KD-values are for monomeric pMHC-TCR interaction as measured by SPR.
Dextramers have stronger signals, less background staining
Ficoll-purified human peripheral blood mononuclear cells (HPBMC) were stained
with Dextramer™, Tetramer and Pentamer according to each product’s recommended
procedures. Gating strategy:
CD4-, CD14- and CD3+. As may be seen, MHC Dextramer™ reagents provide the
highest resolution yet has the lowest background staining.
Dextramers stain 10x stronger
(The data was kindly provided by David Lissauer, Karen Piper, Professor Mark
Kilby and Professor Paul Moss, Birmingham University, Birmingham, UK)
An approximately 10-fold brighter staining of a human T cell clone specific for
the HLA-A2 restricted HY derived epitope FIDSYICQV is observed when stained with
HLA-A2(FIDSYICQV)/PE Dextramer™ than with HLA-A2(FIDSYICQV)/PE Tetramer.
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