Figure 2.

ESO_119-143-specific CD4⁺ T cells can be generated from conventional central memory but not effector memory CD4⁺ T cells or Treg of DR52b⁺ healthy donors. (A) Conventional naïve (N, CD45RA⁺CCR7⁺CD25⁻) and memory (M, CD45RA−CCR7^+/-CD25⁻) cells isolated ex vivo from CD4⁺ T cells of DR52b⁺ healthy donors (n=7), as shown in Online Supplementary Figure S1, were stimulated as in Figure 1 and stained with DR52b/ESO tetramers and anti-CD4 and analyzed by flow cytometry. Dot plots for one donor and data for all donors assessed are shown. (B) Conventional naïve (N, CD45RA⁺CCR7⁺CD25⁻) as well as central memory (CM, CD45RA⁻CCR7⁺CD25⁻) and effector memory (EM, CD45RA⁻CCR7⁻CD25⁻) cells, isolated ex vivo by cell sorting from CD4⁺ T cells of DR52b⁺ donors (n=9) as shown in Online Supplementary Figure S1, were assessed as in A. Dot plots for one donor and data for all donors assessed are shown. (C) NnTreg (CD45RA⁺CCR7⁺CD25⁺CD127^low) and Mtreg(CD45RA⁻CCR7^+/- CD25⁺CD127^low) populations were isolated ex vivo by cell sorting from CD4⁺ T cells of DR52b⁺ donors (n=9), as shown in Online Supplementary Figure S1, and assessed as in A. Dot plots for one donor and data for all donors assessed are shown. Statistical analyses were performed using the Wilcoxon’s matched pairs test. *P<0.05; **P<0.01; ***P< 0.001.