Figure 5

Expression of CXCR4 (CD184) on Slan-DCs and numbers of NK cells and NK-like T cells in HSC grafts collected after mobilization with G-CSF and plerixafor (MZ). Panel A shows the results of a representative experiment aimed at enumerating 6-sulfo-LacNAc⁺ (Slan)-DCs in donors’ PB. Slan-DCs were consistently CD16⁺ and expressed low-level CD14. Panels B and C: PB samples from PMs given G-CSF and single-dose MZ were double-stained with anti-CD184 and anti-M-DC8 mAbs to quantitate CXCR4 expression on Slan-DCs. The Mean Fluorescence Intensity (MFI) of CD184 expression on peripheral CD14⁺ monocytes is shown in a representative experiment out of 4 with similar results. The marker was set according to the proper isotypic control. Panel C shows preserved levels of CD184 on Slan-DCs after HSC mobilization with G-CSF and MZ. This is in contrast with the marked down-regulation of CD184 on monocytes that is depicted in panel B. One representative experiment out of 4 with similar results is shown. Markers were set according to proper isotypic controls. Panels D-G: Frequency and absolute numbers of NK cells and NK-like T cells in TCR-αβ/CD19-depleted haploidentical grafts. After the immunomagnetic removal of TCR-αβ⁺ T cells and CD19⁺ B cells, mononuclear cells contained within the graft were extensively characterized using mAbs directed against CD3, CD16 and CD56 with the aim at identifying NK cells and NK-like T cells. NK cells and NK-like T cells were also quantitated in aliquots of 10 normal BM samples collected under general anesthesia and intended for HLA-identical sibling HSCT. Comparisons among data sets were performed with the Mann Whitney U test for unpaired determinations or with the analysis of variance (ANOVA), as appropriate. HC = healthy control; WBC = white blood cells.