Fig. 7
HlaH35A improves the immunogenicity and protective efficacy of carried antigens by activating ADAM10-Notch signaling in cDCs. a Experimental design for the Fl-BMDC transfer model. Each mouse was intramuscularly injected with 5 × 105 CD11c+MHCIIhi cells. b Serum PA0833-specific IgG levels in mice following two adoptive transfers of Fl-BMDCs treated for 24 h with PBS, PA0833, HPF, HPF with GI254023X, or HPF with DAPT (n = 8–9 per group). The data were pooled from two independent experiments. c Serum PA0833-specific IgG1 or IgG2a levels in mice following two adoptive transfers of Fl-BMDCs treated for 24 h with PBS, PA0833, HPF, HPF with GI254023X, or HPF with DAPT (n = 8–9 per group). The data were pooled from two independent experiments. d Bacterial loads in the lungs determined 48 h after challenge with a sublethal dose (3 × 106 CFU) of PAO1 in mice following two adoptive transfers of Fl-BMDCs treated for 24 h with PBS, PA0833, HPF, HPF with GI254023X, or HPF with DAPT (n = 8–9 per group). The data were pooled from two independent experiments. e Body weight was measured 7 d after challenge with a sublethal dose (3 × 106 CFU) of PAO1 in mice following two adoptive transfers of Fl-BMDCs treated for 24 h with PBS, PA0833, HPF, HPF with GI254023X, or HPF with DAPT (n = 7 per group). f Representative H&E staining of lung sections obtained 48 h after challenge with a sublethal dose (3 × 106 CFU) of PAO1 from mice following two adoptive transfers of Fl-BMDCs treated for 24 h with PBS, PA0833, HPFs with GI254023X, or HPFs with DAPT (n = 3 per group). g TNF-α, IL-1β, IL-6, and IL-10 levels in the lungs were determined 48 h after challenge with a sublethal dose (3 × 106 CFU) of PAO1 in mice following two adoptive transfers of Fl-BMDCs treated for 24 h with PBS, PA0833, HPF, GI254023X, or HPF with DAPT (n = 8–9 per group). The data were pooled from two independent experiments. h Kaplan–Meier survival curves of mice challenged with a lethal dose (1 × 107 CFU) of PAO1, measured 14 days post-infection, following two adoptive transfers of Fl-BMDCs treated for 24 h with PBS, PA0833, HPF, HPF with GI254023X, or HPF with DAPT (n = 10 per group). i Schematic diagram of the proposed mechanism by which HlaH35A enhances immunoprotection via ADAM10 on the surface of DCs. Each data point indicates a biological replicate in (b–d and g). The data are presented as the means ± s.e.m.s. Statistical significance was tested via one-way ANOVA followed by Tukey’s multiple comparisons test in (b–d and g)
