General gating strategy for PBMC samples. Left to right, a debris exclusion gate, a time gate to exclude electronic noise, a singlet gate to exclude doublets and a viability gate to exclude dead cells. T cells (CD3 + SSC-A-low) were gated for CD45RA CD45RO + effector and memory subsets: CD8 + T EM (CD3 + CD8 + CD45RA CD45RO + ), CD4 + T EM (CD3 + CD4 + FOXP3 CD45RA CD45RO + ) and T reg (CD3 + CD4 + FOXP3 + CD45RA – CD45RO + ) cells.
