CXCR4 dysfunction does not alter pDC function. (A) IFN-α secretion (left) by splenic pDC from +/1013 and WT mice stimulated with CpG-ODN (CpG) or not (−) for 18 hours. pDC numbers (right) at the end of the culture. Mean ± SEM, n = 5-6 per group, cumulative data from 2 experiments. (B) Cleaved Caspase 3 levels in freshly isolated spleen pDCs (live CD11c^intCD317⁺ cells) from +/1013 and WT mice. Mean ± SEM, n = 9 per group, cumulative data from 3 experiments. (C) IFN-α secretion by Flt3-l-derived pDCs from +/1013 and WT mice stimulated with CpG-ODN for 18 hours. Mean ± SEM, n = 2 per group, cumulative data from 2 experiments. (D-J) +/1013 and WT mice were injected with CpG-DOTAP (CpG) or DOTAP only (vehicle [veh]). (D) Representative dot plot of the CD45⁺ cells recovered in the peritoneal lavage 24 hours after treatment. (E) Fold increase in the number of cells recovered in the peritoneal lavage, the reference being the WT veh mice. (F) Number of pDCs (live CD45⁺CD11b⁻F4/80⁻CD11c^loLy6C⁺CD317⁺B220⁺ cells), resident macrophages (live CD45⁺CD11b^hiF4/80^hiLy6G⁻Ly6C^{int to hi} cells), and inflammatory myeloid cells (live Infl. myeloid, CD45⁺B220⁻CD11b⁺F4/80^{lo to +}Ly6C^hi cells) in the peritoneal lavage. (G) Serum cytokine levels at different timepoints. (H-I) Surface expression of MHC2 (H) and CD86 (I) by splenic pDC (CD11c^intCD317⁺B220⁺). Representative histograms (left) and the fold increase in the MFI relative to WT veh mice (right) are shown. The gray histogram corresponds to isotype control. (J) Surface expression of CD69 by splenic NK cells (live CD3⁻NK1.1⁺CD49b⁺ cells). Representative dot plots (left), the percentage of CD69⁺ NK cells (middle), and CD69 MFI (right) are shown.) Mean ± SEM, n = 5-6 per group, cumulative data from 3 experiments (D-J). Mice had a C57BL/6J genetic background. Statistical analysis was performed using the 2-tailed unpaired Mann-Whitney test to compare veh- and CpG-treated mice (^#P < .05) or the WT and +/1013 groups (*P < .05).