Figure 6.

Phagocytic processing and migration capacities of blood-derived langerin⁺ DCs. Lethally irradiated F1 (C57BL/6 I-A^b−/−/BALB/c I-E^d+) mice were reconstituted with BM cells isolated from donor Langerin-EGFP CD45.1⁺ C57BL/6 I-A^b+ mice. (A, top) Overlaid histograms show YAe staining on gated host LCs (CD45.2⁺CD11c⁺Langerin-EGFP⁻), donor langerin⁺ dermal DCs (CD45.1CD11c⁺Langerin-EGFP⁺), and donor langerin⁻ dermal DCs (CD45.2⁺CD11c⁺Langerin-EGFP⁻). LCs isolated from F1 (C57BL/6 I-A^b+/+/BALB/c I-E^d+) mice were used as controls. (A, bottom) YAe staining is shown for host LC-derived DCs (CD45.2⁺CD11c⁺Langerin-EGFP⁻), donor langerin⁺ DCs (CD45.1⁺CD11c⁺CD8a⁻Langerin-EGFP⁺), and donor langerin⁻ DCs (CD45.2⁺CD11c⁺CD8⁻langerin⁻) in the skin DLNs. Corresponding YAe isotype control (blue) and WT F1-positive control (green) are shown. (B) Mean values (± the SD) of the percentage of YAe⁺ cells among donor langerin⁺ or langerin⁻ DCs in the dermis and skin DLNs. Representative data from two independent experiments are shown. Each experiment included at least three separately analyzed mice; error bars represent the SD between the results obtained from each of the three mice. (C) Host LCs in the epidermis, donor langerin⁺, or langerin⁻ DCs in the dermis, LC-derived DCs, or donor langerin⁺ or langerin⁻ DCs in the skin DLNs were purified by cell sorter and stained on cytospin for YAe and langerin as described in Materials and methods. Insets show YAe isotype control for each tested DC population. Dashed white lines indicate where cells were spliced from different fields to present more cells within a panel. Bar, 10 μm.