Fig. 5.
Dasatinib allows detection of autoreactive CTL. A. CTL clone IE6, specific for the HLA A2-restricted epitope PPI15-24, was activated with either CMV pp65495-503 or PPI15-24 peptide for 6 h at 37 °C and then assayed for TNFα production by intracellular cytokine staining as detailed in the Materials and methods. B. Staining of CTL clone IE6 with either an irrelevant or HLA A2/PPI15-24 tetramer ± pre-treatment with 50 nM dasatinib for 30 min at 37 °C. C. Representative stainings with HLA A2/PPI tetramer ± pre-treatment with 50 nM dasatinib for 30 min at 37 °C. Left panels: control subject PBMCs; middle panels: type I diabetic patient PBMCs; right panels: a short-term line expanded by one round of peptide stimulation from a type 1 diabetic patient.