NY-ESO-1 edited lymphocytes express high TCR levels and display a preferential stem memory and central memory phenotype. (A) Distribution of T-cell subpopulations (T_SCM, T_CM, effector memory [T_EM], and terminal effector [T_EMRA]) within mock-transduced (mock-trd) and NY-ESO-1–redirected T cells: TR, SE, and CE. The analysis was performed by FACS, and the attribution to the 4 subgroups was made according to the expression of CD45RA, CD62L, and CD95 surface markers, as previously described.³⁴  Histograms related to CD4^pos and CD8^pos T cells are shown. (B) Representative plots of CD45RA, CD62L, and CD95 expression on TCR redirected and mock-transduced T cells. PBMCs are shown as internal control. CD4 and CD8 subsets are separately shown. (C) Percentages of IL-7Rα (CD127) positive cells detected by FACS within CD3^pos T cells and in separately gated CD4^pos and CD8^pos T cells. (D) Percentages of CD27- and/or CD28-expressing cells quantified by FACS and gated on CD3^pos cells. Means and SEM of at least 4 healthy donors are presented. All data were acquired 4 to 5 weeks after S1. TR and SE cells were sorted by NY-ESO-1_157-165 dextramer before the analysis, as described in “Methods.” NY-ESO-1–redirected T cells were expanded by rapid expansion protocol, as previously described,²⁴  and subsequently tested by flow cytometry for the expression of the NY-ESO-1_157-165 Vβ13.1 chain (E) and for the specific binding to a NY-ESO-1_157-165 dextramer (F). Both histograms represent the relative fluorescent intensity (RFI) calculated as follows: (mean fluorescent intensity of TR, SE, and CE cells)/( mean fluorescent intensity of mock-trd cells). Analyses were performed on total CD3^pos cells and on separately gated CD4^pos and CD8^pos cells. Means and SEM of 3 T-cell donors are presented. *P < .05 by paired Student t test.