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OT-1 polarizations were carried out as previously described (Carrio et al, 2004). Briefly, splenocytes from OT-1 mice were cultured at 1×10⁶ cells/mL in 24 well-plates of complete RPMI (UCSF Media Core facility) supplemented with 10% FBS (Omega Scientific, Tarzana, California), 100U/mL penicillin-streptomycin (Fisher Scientific, Hampton, New Hampshire), 2mM L-glutamine (Sigma-Aldrich, St. Louis, Missouri) and 50μM β-mecaptoethanol (Thermo Fisher Scientific, Waltham, Massachusetts ) and 10mM HEPES (UCSF Media Core Facility) in the presence of OVA_257–264 peptide (0.1nM) (Invivogen, San Diego, California) and IL-2 (100U/ml) (Teceleukin) kindly provided by NCI Frederick. After 3 days in culture, activated cells were washed 3 times with RPMI 1640 and recultured in T25 culture flasks at 1×10⁵ cells/mL in the presence of either IL-7, IL-15 (Biolegend, San Diego, California) or IL-2 (Teceleukin) kindly provided by NCI Frederick. (all cytokines 10ng/ml) After 2 additional days in culture, cells were passaged and recultured under the same conditions without peptide for an additional two days for total of 7 days in culture. Viability was confirmed by trypan blue exclusion (Thermo Fisher, Waltham, Massachusetts) or mass cytometry as described below.