Figure 1.

Gating strategy for CD4⁺ and CD8⁺ T cell differentiation. (A,G) Gating of CD4⁺ and CD8⁺ T cells. (B,H) Differential expression of CD45RA and CCR7 defines 6 subsets on CD4⁺ and CD8⁺ T cells: Naïve (CD45RA^hiCCR7⁺), transitional central memory (T-CM) (CD45RA^lo-intCCR7⁺), central memory (CM) (CD45RA⁻CCR7⁺), effector memory (EM) (CD45RA⁻CCR7⁻), late effector memory (L-EM) (CD45RA^lo-intCCR7⁻), and T_EMRA (CD45RA^hiCCR7⁻). CD45RA^hi (Naïve and T_EMRA), and CD45RA^lo-int(T-CM and L-EM) were gated on CD8⁺ T cells (H) and subsequently applied to CD4⁺ T cells (B). CCR7^+/− and CD45RA⁻ (CM and EM) were gated by use of matched isotype control. (C,I) Matched isotype controls for CCR7 and CD45RA on CD4⁺ and CD8⁺ T cells. (D,J) Differential expression of CD25 and CD127 defines two CD25⁺ populations on CD4⁺ and CD8⁺ T cells: CD127⁺CD25⁺ cells and T_Reg (CD127^loCD25^hi) cells. (E,K) Matched isotype controls for CD25 and CD127 on CD4⁺ and CD8⁺ T cells. (F) The CD127⁺CD25⁺ gate is applied to the six CD4⁺ T cell subsets. Frequency of CD127⁺CD25⁺ naïve, T-CM, CM, EM, L-EM, and T_EMRA cells in relation to CD4⁺ T cells. (L) The CD127⁺CD25⁺ gate is applied to the six CD8⁺ T cell subsets. Frequency of CD127⁺CD25⁺ naïve, T-CM, CM, EM, L-EM, and T_EMRA cells in relation to CD8⁺ T cells.