Immunophenotyping of Cryopreserved PBMCs

Cryopreserved PBMC were isolated and used for immunophenotypic staining as previously described (108 (link)). Cells were stained with Zombie NIR fixable viability stain (BioLegend, San Diego, USA) and the following anti-human monoclonal fluorochrome-conjugated antibodies: anti-CD45RA, anti-CD4, anti-TCR-γ/δ (BD Biosciences, Heidelberg, Germany), anti-TCR-Vδ2 (Beckman Coulter Life Sciences, Indianapolis, USA), anti–HLA-DR, anti-CD27, anti-CD279 (PD-1), anti-TIGIT, anti-CD8, anti-CD28, anti-CD39, anti-CD38, anti-CD19, anti-CD3, anti-CD73 and anti-CD14 (all BioLegend) (Supplementary Table 2). Cells were incubated for 30 minutes at room temperature with the respective antibodies. After washing, cells were fixated with 4% paraformaldehyde. All samples were run on a Becton Dickinson LSR Fortessa flow cytometer with FACS Diva version 8 (BD Biosciences).