Figure 3.

IS-SNAs enhances antigen and CpG co-delivery, thereby promoting DC activation and cross-presentation in vivo. (A,B) For co-delivery evaluation, mice were injected subcutaneously with TMR-labeled PSA₆₅₋₇₃ and Cy5-labeled CpG in the form of admix, EM or HM IS-SNAs. Fluorescence signals of CD11c⁺ DCs in dLNs were measured after 2 h by flow cytometry. The uptake (3 mice per group) was shown by representative dot plots (A) and bar graph (B). (C–H) For DC activation and cross-presentation, dLNs (4 mice per group) were harvested and analyzed 24 h after IS-SNA/PSA₆₅₋₇₃ administration. Total number (C) and percentage (D) of CD11c⁺ DCs in dLNs and the expression of co-stimulatory molecule CD40 and CD80 (E) of these DCs were analyzed by flow cytometry. (F–H) Purified DCs from dLNs were co-cultured with PSA₆₅₋₇₃ specific CD8⁺ T cells at 1:2 ratio for 48 h. The supernatant from the co-culture were harvested for analyzing IL-12 (F) and TNF-α (G) production. (H) The DC cross-presentation ability was evaluated using IFN-γ ELISPOT assay. All data are presented as mean ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001, analyzed by two-tailed unpaired Student's t-test.