Figure 2.

Enhanced glyco-dendrimer binding and uptake by moDC via DC-SIGN and primary LC via Langerin. Binding and uptake of G3 (glyco)-dendrimers was evaluated for DC-SIGN⁺ moDC and Langerin⁺ primary LCs. A. Dose-response following a 3 hour pulse, wash and 45 minutes chase of moDC with G3 (glyco)-dendrimers in the presence (dotted line) or absence (solid line) of TLR4 stimulus MPLA. Representative of n=3 measured in triplicate ±SD B. Binding and uptake of (glyco)-dendrimers over-time by moDC following a 45 minutes pulse (no wash) at 4^oC. n=2, ±SD C. Imaging microscopy of moDC following 3 hour incubation at 37 ^oC with glyco-dendrimers (green). Membrane was stained using anti-CD1a (red) and nucleus using DAPI (white) D-E. Involvement of DC-SIGN in binding and uptake of G3 (glyco)- dendrimers was evaluated using a 3 hour pre-incubation with anti-DC-SIGN (C) or 30 minutes pre-incubation with the natural ligand mannan (D) followed by 1 hour incubation with (glyco)-dendrimers. (C) n=4, each symbol represents a donor, (D) representative of n=2 measured in triplicate ±SD F. Binding and uptake of (glyco)-dendrimers over-time by primary LC following a 45 minutes pulse (no wash) on 4^oC. Representative of n=2 measured in triplicate G. Langerin involvement in binding and uptake of (glyco)-dendrimers by primary LCs was evaluated using 30 minutes pre-incubation an anti-Langerin blocking antibody followed by 1 hour co-incubation with (glyco)-dendrimers. Representative of n=3 measured in triplicate ±SD H. Imaging microscopy of primary LC following 3 hours incubation at 37 ^oC with glyco-dendrimers (green). Membrane was stained using anti-CD1a (red) (Statistical analysis: A,E two-way ANOVA Sidak's post hoc; B,D,F two-way ANOVA Tukey's post hoc; C one-way ANOVA Dunnett's post hoc)