Figure 3.

Main general gating strategies for panels A and B

(A) After time lapse, lymphocyte morphology, single and live events selection, samples are gated to exclude CD14⁺ and CD19⁺ cells (monocytes and B cells). Subsequently, CD3^-CD56⁺ cells are gated for further natural killer (NK) cells analysis using CD56 versus CD16 markers. On the other hand, CD3⁺ cells are gated to discriminate unconventional TCRγδ⁺ (γδT cells), TCRVα7.2⁺ (MAIT cells) and TCRVα2.4⁺ (iNKT cells) lymphocytes. From the remaining T cell population, cells are gated by CD4⁺ versus CD8⁺ cells. NKG2A⁺NKG2C^- (CD8⁺NKG2A⁺) and NKG2A^-NKG2C⁺ (CD8⁺NKG2C⁺) cells are then identified from the CD8⁺ T cell population. Then, the NKG2A^-NKG2C^- CD8⁺ subset as well as the total CD4⁺ cells are gated according to differentiation markers; CD45⁺CCR7⁺ T naïve (TN) cells, CD45^-CCR7⁺ central memory (CM) cells, CD45RA^-CCR7^- effector memory (EM) cells, and CD45RA⁺CCR7^- terminally differentiated (TEMRA) cells.

(B) Correspondingly, example graphs showing manually gated major populations (bottom figures) as proportions from their parent CD3⁺ T cell population (left), CD4⁺CD8^- and CD4^-CD8⁺ T cells (center), and NK cells (right).Bar graphs represent the means; error bars represent the standard deviation; and dots represent individual donors (N=10). One-way ANOVA with Tukey’s multiple comparison test. Significance is indicated as ∗∗∗∗P ≤ 0.0001, ∗∗∗P ≤ 0.001, ∗∗P ≤ 0.01, ∗P ≤ 0.05, or P > 0.05 (not significant (ns)).