Antibodies, tetramers, and flow cytometry

All antibodies used for flow cytometry were purchased from either ThermoFisher, BD Biosciences, or BioLegend. The following antibodies were used to discriminate cell surface or intracellular phenotype: TCRβ (H57-597), CD3 (500A2), CD90.2 (53-2.1), CD4 (RM4-5), CD8b (H35-17.2), CD45.1 (A20), CD45.2 (104) CD44 (IM7), IFN-γ (XMG1.2), TNFα (MP6-XT22), IL-17α (eBio17B7), IL-2Rα/CD25 (PC61), Vα2 (B20.1) and Foxp3 (FJK-16S). Dead cells were discriminated in all experiments using LIVE/DEAD fixable dead stain (ThermoFisher). All stains were carried out in media containing anti-CD16/32 blocking antibody (clone 93, ThermoFisher). For intracellular cytokine staining, cells were fixed in BD Cytofix buffer (BD Biosciences) and stained in BD PermWash buffer (BD Biosciences). For Foxp3 staining, cells were fixed and permabilized using the FoxP3/Transcription factor staining buffer set according to the manufacturer’s directions (ThermoFisher). APC-conjugated CBir1_464-72 tetramers (YSNANILSQ) and PE-conjugated HH1713_172-86 and HH1713_230-44 tetramers (QESPRIAAAYTIKGA and GNAYISVLAHYGKNG, respectively) were provided by the NIH Tetramer Core Facility (Atlanta, GA). All tetramer stains were performed at room temperature for 45–60 minutes. All flow cytometry was acquired on an LSRFortessa FACS analyzer, and cell sorting was carried out on a FACS Aria (BD Biosciences).