Fig. 1

Gating strategy for the identification of human T_SCM. T cells were identified by gating on lymphocytes (SSC versus FSC), singlets (FSC-H versus FSC-A) and live T cells (SSC versus LIVE/DEAD). CD4⁺ and CD8⁺ T cells were simultaneously identified with anti-CD4 and anti-CD8 antibodies. a Gating strategy of fresh blood cells. After gating on CD4⁺ and CD8⁺ cells, T_CM and T_EM subpopulations were identified based on CCR7 and CD45RO expression. In the gated CCR7⁺CD45RO⁻ population, cells expressing CD45RA and CD27 were further analyzed. In this later population (CCR7⁺CD45RO⁻CD45RA⁺CD27⁺), T_N and T_SCM were identified based on the CD95 expression. T_N is defined as CCR7⁺CD45RO⁻CD45RA⁺CD27⁺CD95⁻ whereas T_SCM subpopulation is defined as CCR7⁺CD45RO⁻CD45RA⁺CD27⁺CD95⁺. Red arrows indicate the sequential gating strategy. b Gating strategy of 10 days culture cells. After gating on CD4⁺ and CD8⁺ cells, T_CM and T_EM subpopulations were identified based on CCR7 and CD45RO expression. In the gated CCR7⁺CD45RO⁻ population, cells expressing CD45RA and CD27 were further analyzed. In this later population (CCR7⁺CD45RO⁻CD45RA⁺CD27⁺), T_SCM were identified based on the CD95 expression. T_SCM subpopulation is defined as CCR7⁺CD45RO⁻CD45RA⁺CD27⁺CD95⁺. Similarly, in the gated CCR7⁺CD45RO⁺ population, cells expressing CD45RA, CD27 and CD95⁺ identify a T_SCM-like subpopulation, which is defined as CCR7⁺CD45RO⁺CD45RA⁺CD27⁺CD95⁺. Red arrows indicate the sequential gating strategy