Anti cd8 bv650

Lymphocytes from human thymi, cord blood, lymph nodes (mesenteric and pancreatic), spleen and blood samples were cell surface stained with anti‐CD3‐AF700 (557943)/anti‐CD3‐PE‐CF594 (562280), anti‐CD19‐BV570 (BioLegend, San Diego, CA, USA; 302236), anti‐CD4‐APC‐H7 (560158)/anti‐CD4‐BV650 (563875), anti‐CD8‐BV421 (BioLegend, 301035)/anti‐CD8‐BV650 (563875)/anti‐CD8‐PerCP‐Cy5.5 (565310), anti‐CD27‐BV711 (563167), anti‐CD45RA‐FITC (555488), anti‐PD‐1‐PE‐Cy7 (561272) antibodies (all from BD Biosciences, San Jose, CA, USA), unless otherwise stated with their catalogue number in brackets) and LIVE/DEAD (Aqua, Near‐IR, Molecular Probes, Eugene, OR, USA) or 7‐amino‐actinomycin D (Sigma‐Aldrich, Darmstadt, Germany) on ice for 30 min in PBS. Human PBMCs were sorted for CD27⁺CD45RA⁺ naïve or CD27⁺CD45RA⁻ T_CM CD4⁺ and CD8⁺ T cell populations using FACS Aria III (BD Biosciences). Alternatively, cells were fixed and permeablized using the eBioscience Foxp3 staining kit (ThermoFisher Scientific, Waltham, MA, USA) then intracellularly stained with anti‐SATB1‐AF647 (BD Biosciences, 562378) and anti‐GzmB‐AF700 (BD Biosciences, 560213). Samples were acquired using a BD LSR Fortessa (BD Bioscience) and analyzed using FlowJo software (V10.4.2 and 10.5, Treestar, Ashland, OR, USA) and GraphPad Prism (GraphPad Software, La Jolla, CA, USA, V7.02).

Zombie UV™ Fixable Viability Kit, anti-CD45 BV570, anti-CD3 BV605, anti-CD4 BV421, anti-CD8 BV650, anti-CD4 APC-Fire750, anti-CD73 PE/Dazzle 594, anti-CD56 PE, anti-CD14 PerCP/Cy5.5, anti-CD16 Alex Fluor 700, anti-CD19 APC-Fire750, anti-HLA-DR PE-Cy5, anti-PD-1 BV421, anti-CD16 PE, anti-IFNγ A700, anti-IL-13 APC, anti-IL-5 APC, anti-TNFα Alexa Fluor 488, and anti-IL-21 PE were purchased from BioLegend (London, UK). Anti-CD25 BB515, anti-CD4 BUV496, anti-CD39 BV711, anti-Perforin Alexa Fluor 647, anti-Granzyme B Alexa Fluor 700, anti-Ki-67 Alexa Fluor 700, anti-CD45RA PE, anti-IL-2 PerCP/Cy5.5, and Fixation/Permeabilization Solution Kit were purchased from Becton Dickinson. Anti-FOXP3 APC, anti-CD56 APC-eFluor780, and eBioscience™ (San Diego, CA, USA) Foxp3/Transcription Factor Staining Buffer Set were purchased from eBioscience TermoFisher scientific. Anti-TIM-3 APC was purchased from R&D systems. Anti-CD137 PE and FcR blocking reagent were purchased from Miltenyi (Bergisch Gladbach, Germany).
Peptides for the PSA, PSMA, PSCA, and PAP pools (Supplementary Table S1) were synthetized and lyophilized by the Peptide and Tetramer Core Facility of the Department of Oncology at UNIL-CHUV (Lausanne, Switzerland), and 5T4 pools (Supplementary Tables S2 and S3) were synthetized and lyophilized by JPT Peptide Technologies.