Fig. 2: AECM@PC7A stimulates APC maturation and T-cell priming in vitro and in vivo.

a–d Scheme showing determination of APC maturation and T-cell priming in vitro (a). BMDCs were treated with CMs or CMs-derived nanovaccines at a final concentration of 20 μg/mL for 18 h, expression of CD80 and CD86 on BMDCs were evaluated by flow cytometry (b, n = 2 independent experiments for No adjuvant groups, n = 3 independent experiments for PC7A groups). The treated BMDCs were harvested and co-cultured with CellTrace^TM Violet-labelled OT-I splenocytes for 48 h, CD8⁺ T-cell proliferation was shown by representative FACS histograms (c) and quantification (d, n = 3 independent experiments/group). e–k Vaccination timeline in mice (e). C57BL/6 mice received s.c. vaccinations on days 0, 7 and 14, and peripheral blood and lymph nodes were collected at day-21 for analysis. Representative scatter plots (f) and quantification (g, n = 4 mice/group) showing CD86 and CD80 expression on DCs from lymph nodes. Representative scatter plots (h) and quantification (i, n = 4 mice/group) showing OVA tetramer⁺ CD8⁺ cells in PBMCs. Representative scatter plots (j) and quantification (k, n = 4 mice/group) showing IFN-γ producing CD8⁺ cells in PBMCs upon OVA_257-264-peptide restimulation. l, WT or Baft3^-/- mice were s.c. vaccinated with AECM@PC7A three times at a dose of 300 μg per shot, PBMCs were collected at day-21 and re-stimulated with OVA_257-264 peptide, the IFN-γ secretion in CD8⁺ T-cells were determined by ICS (n = 3 mice/group). m C57BL/6 mice received AECM@PC7A or PBS s.c vaccinations together with Clodronate- or control- liposomes i.p. treatment as indicated, PBMCs were collected at day-21 and re-stimulated with OVA_257-264 peptide, the IFN-γ secretion by CD8⁺ T-cells were determined by ICS (n = 3 mice/group). In (b, d, g, i, k–m), representative data from three independent experiments are presented as means ± s.e.m. Statistical significance was calculated by ordinary one-way ANOVA (b, d, g, I, k) and Student’s two-sided unpaired t-test (l, m). Source data are provided as a Source Data file. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. NS, not significant. Source data are provided as a Source Data file.