Fig. 3

Progressive expression of KLRs and GPR56 is associated with cytokine production during CD4+ memory T-cell development. a Association of surface marker expression with cytokine expression in CD4⁺ T cells from peripheral blood of healthy individuals shown as exemplary dot plots and summarizing box and whisker plots (% of cytokine expressing cells within total or marker positive subset, whiskers extend to the minimum and maximum) of n = 5. Cells were restimulated with PMA and Ionomycin. b Representative t-SNE plots showing surface marker and cytokine expression pattern of pre-gated CD4⁺ T cells (excluding CD25^highCD127^low Treg cells) upon short-term phorbol 12-myristate 13-acetate (PMA)/Ionomycin stimulation. The highlighted area marks CD4⁺ T_EM and T_EMRA cells identified by the absence of CCR7 and the expression pattern of CD45RA. c Wanderlust analysis based on the trajectory of CD45RA and CCR7. Relative median surface marker and intracellular cytokine expression within CD4⁺ T cells from blood of five healthy individuals upon short-term PMA/Ionomycin stimulation are shown as described within Methods. P-values were determined by non-parametric matched-pairs Friedman’s test with post-hoc Dunn’s multiple comparison test. *p < 0.05, **p < 0.01