Recombinant human il 2 or mouse il 15

LN T cells from WT and Ptpn22^−/− were cultured in RPMI 1640 media (Invitrogen) supplemented with 10% FCS, L-glutamine, antibiotics and 50μM 2-mercaptoethanol. N4, T4, G4 and RTYTYEKL peptides (Peptide Synthetics) were added to culture media at concentrations stated in figure legends. For growth of CTLs, OT-1 cells were stimulated for 2d with 10⁻⁸M N4, washed, then expanded/differentiated in 20 ng/ml recombinant human IL-2 or mouse IL-15 (Peprotech) for a further 4 d. For cytokine recall responses, cells were re-stimulated in the presence of 2.5 μg/ml brefeldin A (Sigma) for 4h, prior to fixation and intracellular staining. For analysis of glucose uptake, following 24h peptide stimulation, cells were incubated for 45 min with 10 μM NBDG, then washed twice in PBS prior to fixation in 2% paraformaldehyde and FACS analysis. Relative glucose uptake was calculated as the mean fluorescence intensity of NBDG staining following stimulation, with background fluorescence subtracted.