Figure 2. Detailed analysis of targeted DC subsets.

1μg IgG2a control or anti-DEC-205 mAb were injected intradermally into the ears of C57BL/6 mice. The injection site was immediately treated with imiquimod cream, or left untreated. At the indicated time points, targeting antibody was detected on permeabilised lymph nodes cells by anti-rat IgG / APC, and the different subset of DCs were distinguished by staining for CD11c, CD11b, CCR7, CD8α and Langerin. (A, upper panel) Gating strategy for CD11c⁺ CCR7^neg CD8α^neg CD11b-/⁺ LN DCs, CD11c⁺ CCR7^neg CD8α⁺ CD11b^low LN DCs, CD11c⁺ CCR7⁺ CD8α^neg Langerin^neg CD11b⁺ dermal DCs and CD11c⁺ CCR7⁺ CD8α^neg Langerin⁺ CD11b⁺ LCs / dermal DCs. (A, lower panel) Detection of DEC-205 targeting antibody (empty histogram) and IgG2a isotype control (filled histogram) on four different DC subsets. Results are representative of two independent experiments. (B) Percentage of CD11b⁺ cells among the four different DC subsets, irrespective of targeting. (C) Relative proportions of each DC subset in untreated mice or mice treated with imiquimod cream. Data from (B) and (C) are pooled from two independent experiments, with at least 2 mice per condition. Significance values apply to the comparison of identical populations (i.e., same color columns) with or without imiquimod treatment.