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Protein kinase inhibitors substantially improve the physical detection of T-cells with peptide-MHC tetramers
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Lissina, Anna, Ladell, Kristin, Skowera, Ania, Clement, Matthew, Edwards, Emily, Seggewiss, Ruth, Berg, Hugo van den, 1968-, Gostick, Emma, Gallagher, Kathleen, Jones, Emma, Melenhorst, J. Joseph, Godkin, Andrew J., Peakman, Mark, Price, David A., Sewell, Andrew K. and Wooldridge, Linda. (2009) Protein kinase inhibitors substantially improve the physical detection of T-cells with peptide-MHC tetramers. Journal of Immunological Methods, Vol.340 (No.1). pp. 11-24. ISSN 0022-1759
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Official URL: http://dx.doi.org/10.1016/j.jim.2008.09.014
Abstract
Flow cytometry with fluorochrome-conjugated peptide-major histocompatibility complex (pMHC) tetramers has transformed the study of antigen-specific T-cells by enabling their visualization, enumeration, phenotypic characterization and isolation from ex vivo samples. Here, we demonstrate that the reversible protein kinase inhibitor (PKI) dasatinib improves the staining intensity of human (CD8+ and CD4+) and murine T-cells without concomitant increases in background staining. Dasatinib enhances the capture of cognate pMHC tetramers from solution and produces higher intensity staining at lower pMHC concentrations. Furthermore. dasatinib reduces pMHC tetramer-induced cell death and substantially lowers the T-cell receptor (TCR)/pMHC interaction affinity threshold required for cell staining. Accordingly, dasatinib permits the identification of T-cells with very low affinity TCR/pMHC interactions, such as those that typically predominate in tumour-specific responses and autoimmune conditions that are not amenable to detection by current technology. (c) 2008 Elsevier B.V. All rights reserved.
| Item Type: | Journal Article |
|---|---|
| Subjects: | Q Science > QD Chemistry Q Science > QR Microbiology > QR180 Immunology |
| Divisions: | Faculty of Science > Mathematics Faculty of Science > Centre for Systems Biology |
| Journal or Publication Title: | Journal of Immunological Methods |
| Publisher: | Elsevier BV |
| ISSN: | 0022-1759 |
| Date: | 1 January 2009 |
| Volume: | Vol.340 |
| Number: | No.1 |
| Number of Pages: | 14 |
| Page Range: | pp. 11-24 |
| Identification Number: | 10.1016/j.jim.2008.09.014 |
| Status: | Peer Reviewed |
| Publication Status: | Published |
| Access rights to Published version: | Restricted or Subscription Access |
| Funder: | Cardiff University Link Chair scheme, TheWellcome Trust, Juvenile Diabetes Research Foundation, Department of Health via the National Institute for Health Research (NIHR) Comprehensive Biomedical Research Centre award |
| Grant number: | 7-2005-877 |
| URI: | http://wrap.warwick.ac.uk/id/eprint/28583 |
Data sourced from Thomson Reuters' Web of Knowledge
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