Protein kinase inhibitors substantially improve the physical detection of T-cells with peptide-MHC tetramers

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• 作者：Anna Lissina ; Kristin Ladell ; Ania Skowera ; Matthew Clement ; Emily Edwards ; Ruth Seggewiss ; Hugo A. van den Berg ; Emma Gostick ; Kathleen Gallagher ; Emma Jones ; J. Joseph Melenhorst ; Andrew J. Godkin
• 关键词：T cell ; Tetramer ; Low avidity CTL ; Cancer immunology ; Autoimmunity
• 刊名：Journal of Immunological Methods
• 出版年：2009
• 出版时间：1 January 2009
• 年：2009
• 卷：340
• 期：1
• 页码：11-24
• 全文大小：2524 K

文摘

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.