Fig. 3
Characterization of CD45RA-depleted DLIs. Apheresis product was either processed immediately after apheresis (fresh mob LP, red) or after storage at 2–6 °C for 48 h (aged mob LP, patterned red). CD45RA-depleted DLI product (fresh TCB1, green) was analyzed immediately after processing of fresh or aged mob LP or after storage at 2–6 °C for 48 h (aged TCB1, light green). A Relative contribution of individual mature leukocyte subpopulations in starting material and target cell fractions among CD45+ leukocytes at the different time-points. The percentage of each subtype was normalized to 100% CD45+ leukocytes. B Percentage of viable CD45+7-AAD− leukocytes in all fractions at different time-points. Depletion efficiency of naïve CD45RA+ T cells and B cells (universally CD45RA+) was compared for products (C) derived from mobilized apheresis product (mob LP), processed immediately after apheresis (fresh mob LP) or after storage at 2–6 °C for 48 h (aged mob LP) and D derived from processes using NS or LS depletion reagents, respectively. Log depletion for each individual run is depicted with dots and squares representing products generated from fresh and aged mob LP or from NS and LS reagents, respectively. Red line indicates mean values achieving mean log depletions of around 4 log in all tested settings. Box plots and whiskers represent median with interquartile range and min and max values of all 10 depletion runs, respectively. Mean yield of CD34+ stem cells, NK cells, TCRγδ+ cells and CD3+ T cells among total CD45+ cells in depleted products (E) derived from fresh or aged mob LP and F from processes using NS or LS depletion reagents showing a high loss of cells of interest. G The CD4/CD8 ratio among total T cells was 6.5-fold increased in all depleted products. H Residual alloreactive CD45RA+ T cells in CD45RA-depleted products were calculated per 1 × 106 CD3+ T cells which are the main active component of a DLI product. All data were tested for statistical significance using Student’s t-test. Shown is mean ± SEM, n = 4–10 individual runs; ns (not significant) indicates a p > 0.05, *: p < 0.05, ***: p < 0.001. P-values are reported above the corresponding groups