Figure 1. Sorting strategy to obtain cellular subsets.

CD4⁺ T cells were negatively selected by immunomagnetic beads from peripheral blood mononuclear cells from 2 HIV-infected donors on ART at 2 time points corresponding to 2 and 9 years after ART initiation .T_N cells were enriched by sorting for CD45RA⁺CCR7⁺CD27⁺ cells. T_CM cells were CD45RA^–CCR7⁺CD27⁺, T_TM cells were CD45RA^–CCR7^–CD27⁺, while T_EM cells were CD45RA^–CCR7^–CD27^–. Finally, CD45RA_dim cells were also collected in an effort to sample almost all CD4⁺ T cell subsets. For selected experiments, we also sorted CD95^– T_N cells (CD45RA⁺CCR7⁺CD27⁺CD95^–) and T_SCM cells (CD45RA⁺CCR7⁺CD27⁺CD95⁺). Purity was determined after sorting by flow cytometry. Here, we show 1 representative experiment. For these sorting experiments, we did not sort CD45RA⁺CD27^–CCR7^– cells (T_EMRA cells; ref. 66). The low purity of T_SCM cells can be explained by the low levels of CD95 expressed by these cells, resulting in limited separation between the CD95⁺ and CD95^– population. T_EMRA, effector memory reexpressing CD45RA.