Fig. 2

Flow cytometric panels for characterization of CD45RA-depleted DLI (TCB1) and TCRαβ/CD19-depleted HSPCs (TCB2) in comparison to apheresis product (mob LP). A The gating strategy using the express mode “TCRab_CD45RA_Depletion_h_02” determines automatically the TCRαβ⁺ and CD45RA⁺ frequencies in the different fractions before and after depletion. Additionally, this panel includes the identification of the CD34⁺ cell population among CD45⁺ leukocytes, however the stem cell enumeration was determined using the BD’s CE-certified SCE kit following stringent ISHAGE gating (panel not shown, [26]). The arrows indicate, left to right, TCRαβ over TCRγδ gated on T cells, CD45RA over CD45RO gated on T cells, and CD45RA over CD45RO gated on TCRαβ T cells. The same analysis is shown, from top to bottom, for apheresis product (Mob LP), CD45RA-depleted DLI and TCRαβ/CD19-depleted HSPCs. CD45RA-depletion depletes almost all TCRαβ T cells, but the inverse is not true for TCRαβ-depletion: the lower panel demonstrates a high frequency of CD45RA⁺ T cells (lower panel, middle) despite almost complete depletion of TCRαβ T cells (lower panel, left). B The gating strategy using the express mode “Immune_Cell_Composition_human” facilitates automatic determination of cell concentration, viability of leukocytes, cell composition and frequency of B cells. The arrows identify B cells. Both CD45RA-depletion (middle panel) and TCRαβ/CD19-depletion (lower panel) almost completely deplete B cells from DLI (middle) or HSPC product (lower)