Immunophenotype characterization of reactive T cells against fungi. (A) t-SNE plots (described in Figure 3A) for panfungal T cells cultured for 6 hours with nonpulsed DCs (DC control) or fungus-pulsed DCs (DC pulsed). Gates show 3 different populations of cells responding to fungi (these populations were only present in panfungal T cells cultured with fungal pulsed DCs). (B) Percentage of the 3 populations of reactive panfungal T cells after 6-hour coculture with fungal lysate–pulsed DCs analyzed by CyTOF. Results are means of 3 experiments (±SEM) and shown as percentage of CD4⁺ T cells. (C) Expression of exhaustion and phenotype markers and cytokines (shown on the y-axis) in the 3 different reactive T-cell populations (shown on the x-axis) of panfungal T-cell products after 6-hour coculture with fungal lysate–pulsed DCs and analyzed by CyTOF. Results are expressed in terms of heat map in which each square represents the mean value percentage of CD4⁺ T cells from 3 experiments. R1, reactive T-cell population 1; R2, reactive T-cell population 2; R2(17), reactive T-cell population 2 (IL-17A⁺); T_cm, central memory T cells (CD45RA ⁻CCR7⁺); T_eff, terminal effector T cells (CD45RA⁺CCR7⁻); T_em, effector memory T cells (CD45RA⁻CCR7⁻); T_n, naive T cells (CD45RA⁺CCR7⁺); T_reg, regulatory T cells (CD137⁺CD154⁻).