Figure 6.

Impact of CMV history on manufacture of untreated and Rapa-TCPs. Untreated (w/o, blue) and Rapa-TCPs (red) of n = 19 patients (9 with so far no recorded CMV viremia; 4 with recent CMV viremia and 6 with a history of CMV viremia)/13 HDs. (A) Yield of TCPs = total cell number derived from 20 ml of patient blood on d21. (B) CD4/CD8 ratio of TCPs on d14. (C,D) Proportions of CD45RA⁻ CCR7 ⁺ T_CM among CD4⁺ (C) and CD8⁺ T-cells (D) in TCPs as determined per gating strategy shown in Figure 5C on d14. (E,F) Proportions of CMV-reactive CD4⁺ (E) and CD8⁺ (F) IFNγ-producers detected by intracellular staining in multicolor flow cytometry after 6 h stimulation with autologous LCLs loaded CMV_IE−1/pp65 peptide pools at a ratio of 1:10 and addition of BFA after 1 h on d21. Gating strategy is shown in Figure 5F. (G) Specific killing of CMV_IE−1/pp65 peptide pool loaded autologous LCLs determined by ratio with unloaded allogenic LCLs at 1:10 ratio with TCPs after incubation overnight. All data were tested for normality with Kolmogorov-Smirnov test; significant differences for paired samples determined with paired t-test if normally distributed or Wilcoxon's matched-pairs signed rank test and for unpaired samples with unpaired t-test if normally distributed or Man Whitney's test. P-values below 0.05 are indicated by * and defined to be significant.