CD8+ Treg cells were induced in an in vitro culture system as previously described (8, 55). Specifically, PBMCs (1 × 106 cells/mL) were stimulated with 0.1 ng/mL anti-CD3 antibody (BioLegend, OKT3, 317315) and 5 ng/mL recombinant human IL-15 protein (Sino Biological, GMP-10360-HNCE) for 6 days. CD8+CCR7+ Treg cells were negatively selected with EasySep human naive CD8+ T cell isolation kits (STEMCELL Technologies, Inc.). Over 90% of the purified cells had a CD8+FOXP3+CCR7+ phenotype.

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