Flow cytometry raw files

<b>Sample preparation and data acquisition</b>Flow cytometry analysis was performed using a Cytek® Aurora full spectrum flow cytometer (Cytek Biosciences, California, USA). Cryopreserved PBMCs were gently thawed in a water bath at 37 °C with a mean recovery of 81.28 % viable cells assessed with the Zombie NIR™ Fixable Viability Kit (BioLegend, San Diego, CA, USA). After incubating 1 × 10⁶ PBMCs in 2.5 µg Fc block for 10 min at room temperature, cells were stained with anti-CD3 (BUV395, clone SK7), anti-CD4 (PerCP, clone SK3), anti-CD8 (BV750, clone SK1), anti-CD16 (PE-Cy7, clone 3G8), anti-CD25 (BUV805, clone M-A251), anti-CD56 (BUV563, clone NCAM16.2), anti-CD20 (APC, clone L27), and anti-CD19 (BV480, clone SJ25C1) antibodies (all from BD Biosciences, NJ, USA). In brief, cells were incubated in the dark with a master mix containing Brilliant Stain buffer (BD Biosciences) and antibodies against surface antigens for 30 min at 4°C. After washing with FACS buffer, the BD Pharmingen™️ Transcription Factor Buffer Set was used, and cells were fixed for 40 min at 4 °C in the dark. Thereafter, intracellular staining was done by incubating cells with an anti-Foxp3 antibody (PE, clone 259D/C7) for 45 min at 4 °C in the dark. After washing, cells were resuspended in FACS buffer and acquired on the flow cytometer within 2 hours after finishing the staining protocol.<b>Data processing</b>Gating was performed using FlowJo™ 10.10.0 (Figure 1C). B cells were phenotyped as CD3^-CD56^-CD19⁺CD20⁺, Natural Killer T (NKT) cells as CD3⁺CD56⁺, Natural Killer (NK) cells either as CD56^brightCD16^-(NK^bright) or CD56^dimCD16⁺ (NK^dim), and regulatory T cells (T_regs) as CD4⁺CD25⁺Foxp3⁺. The person analyzing the samples was blinded to the participants’ group allocation. Analysis of total blood cell counts was performed from EDTA blood using a hematology analyzer (SYSMEX XP-300, Norderstedt, Germany). The lymphocyte count was then used to calculate the absolute number of peripherally circulating lymphocyte subsets according to the cell proportions derived by flow cytometry.