Figure 4

Overview of the gating strategy for panel ONE 03: T cell activation. Expression of CD57 or HLA-DR and loss of CD27 or CD28 expression was used as a sign of T cell activation, as previously described [27–32]. The data file of the stained lysed (EDTA spiked) whole blood (WB) was analyzed as follows: exclusion of non-single events and gating of CD45⁺ leukocytes as shown for panel ONE 01 (Figure 2); gating of CD3⁺ T cells (anti-CD3 versus sideward scatter); and gating of CD4⁺ as well the CD8⁺ T cells (anti-CD4 versus anti-CD8), for both subsets gating on CD57⁺ cells (anti-CD57 versus sideward scatter), HLA-DR⁺/CD45RA⁺ (naive, and HLA-DR⁺/CD45RA^- (memory), and CD27^-/⁺ and CD28^-/⁺ subsets (anti-CD27 versus anti-CD28). WB, whole blood.