REAGENT or RESOURCE	SOURCE	IDENTIFIER
Flow cytometry antibodies/reagents
Anti-human CD3	BioLegend	Cat# 317308 (PE), 317306 (FITC), 317342; RRID AB_571913, AB_571907, AB_2563410
Anti-human CD4	BioLegend	Cat# 317408; RRID AB_571951
Anti-human CD8	BioLegend	Cat# 344722 (APC), 344704 (FITC), 344714; RRID AB_2075388, AB_1877178, AB_2044006
Anti-human CD69	BioLegend	Cat# 310914; RRID AB_314849
Anti-human CD25	BioLegend	Cat# 302606; RRID AB_314276
Anti-human HLA-DR	BioLegend	Cat# 307610; RRID AB_314688
Anti-human/mouse Granzyme B	BioLegend	Cat# 515403; RRID AB_2114575
Anti-human TNF-alpha	BioLegend	Cat# 502908; RRID AB_315260
Anti-human IFN-gamma	BioLegend	Cat# 506510; RRID AB_2623781
FITC Mouse IgG1, κ Isotype Ctrl Antibody	BioLegend	Cat# 400108
PE Mouse IgG1, κ Isotype Ctrl Antibody	BioLegend	Cat# 400112
APC Mouse IgG1, κ Isotype Ctrl Antibody	BioLegend	Cat# 400120
True-Nuclear Transcription Factor Buffer Set	BioLegend	Cat# 424401
SYTOX Blue Viability Dye	Thermo Fisher Scientific	Cat# S34857
LIVE/DEAD Fixable Aqua Dead Cell Dye	Thermo Fisher Scientific	Cat# L-34966
Biological samples
Human blood from healthy donors	UHN-REB (No. 11-0343-CE)	N/A
Chemicals, peptides, and recombinant proteins
Lymphoprep	STEMCELL Technologies	Cat# 07851
RPMI 1640	Wisent	Cat# 350-000-CL
HBSS	Wisent	Cat# 311-513-CL
Human AB Serum	Sigma	Cat# H4522
Fetal Bovine Albumin	Thermo Fisher Scientific	Cat# 12483020
Bovine Serum Albumin	BioShop	Cat# ALB001
Human IL-2	Miltenyi Biotec	Cat# 130-097-743
HEPES (1 M)	Thermo Fisher Scientific	Cat# 15630-080
Penicillin-Streptomycin-Glutamine (100×)	Thermo Fisher Scientific	Cat# 10378-016
Ammonium chloride (NH4Cl)	Sigma	Cat# 213330
Potassium bicarbonate (KHCO3)	Sigma	Cat# 237205
EDTA (0.5 M)	Thermo Fisher Scientific	Cat# 15575-020
Phosphate-buffered saline (PBS)	Thermo Fisher Scientific	Cat# 10010049
HBSS	Thermo Fisher Scientific	Cat# 14025092
5-Aza-2-deoxycytidine	Sigma	Cat# A3656-5MG
Critical commercial assays
CD8+ T cells negative selection kit	Miltenyi Biotec	Cat# 130-096-495
CD8 MicroBeads	Miltenyi Biotec	Cat# 130-045-201
Dynabeads Human T-Activator CD3/CD28	Thermo Fisher Scientific	Cat# 11131D
Cell Stimulation plus Protein secretion inhibitor Cocktail (Cocktail 1 in this protocol)	Thermo Fisher Scientific	Cat# 00-4975-03
Protein secretion inhibitor Cocktail (Cocktail 2 in this protocol)	Thermo Fisher Scientific	Cat# 00-4980-93
QuadroMACS™ Starting Kit (LS)	Miltenyi Biotec	Cat# 130-091-051
Other
DynaMag-2 Magnet	Thermo Fisher Scientific	Cat# 12321D
Suspension, U-bottom, 96-well culture plate	Sarstedt	Cat# 83.3925.500
96-Well Clear V-Bottom TC-Treated Microplate	VWR	Cat# 29442-068
Bovine Serum Albumin	BioShop	Cat# ALB001
10 mL Pipette Reservoirs, Sterile	Diamed	Cat# MTCP8010-5S
UltraComp eBeads™ Plus Compensation Beads	Thermo Fisher Scientific	Cat# 01-3333-41
ArC™ Amine Reactive Compensation Beads	Thermo Fisher Scientific	Cat# A-10346
Trypan Blue Solution, 0.4%	Thermo Fisher Scientific	Cat# 15250061
Software and algorithms
FlowJo	BD Biosciences	www.flowjo.com