Maturation of human DC subsets upon TLR ligand injection in vivo. (A) Fold up-regulation of CD86, CD83, CD274, CD40, and HLA-DR on CD1c⁺ cDCs (top), CD303⁺ pDCs (middle), and CD141⁺ cDCs (bottom). HuNSG mice were injected intraperitoneally with 50 μg/mouse polyICLC, 20 μg/mouse GLA IDC 1001, 25 μg/mouse protamine/RNA, 20 μg/mouse R848, and 50 μg/mouse CpG ODN 2216 or PBS. At 14 hours after injection, splenocytes were isolated and stained for flow cytometry. Fold up-regulation was calculated from the mean fluorescence intensity (MFI) in relation to the mean of the corresponding PBS samples. The graph represents composite data from 5 independent experiments. Each data point represents one individually analyzed mouse. Statistical analysis was performed with the Mann-Whitney U test. (B) Immunohistochemistry for DC markers on spleen sections. HuNSG mice were injected with PBS (top) or with polyICLC (bottom) and euthanized after 14 hours. DEC-205 serves as a DC marker and DC-LAMP and HLA-DR as DC maturation markers.