Fig 1: Sequential gating strategy for identification of T-cell subsets in human PBMCsRepresentative plots showing the gating hierarchy applied to (A) unstimulated and (B) SEB-stimulated PBMCs samples. Singlets were identified using FSC-A versus FSC-H, followed by exclusion of dead cells and dump-positive cells (CD14, CD16, CD19, CD56).Lymphocytes were gated based on FSC/SSC characteristics. CD3+ T-cells were subsequently identified and separated into CD4+ and CD8+ subsets.Regulatory T-cells were defined as CD3+ CD4+ CD25+FoxP3+ cells. Memory and homing phenotypes were further characterized based on CD45RA/CD45RO, CCR7, CLA, and CCR9 expression.