Fig. 7. Albumin-binding lipid-based pulmonary vaccine. (a) Structure of amph-vaccines. Upper: schematic of 1,2-distearoyl-sn-glycero-3phosphoethanolamine (DSPE) conjugated to cysteine(Cys)-terminated peptide via a polyethylene glycol (PEG) spacer. Lower: schematic of diacyl lipid tail conjugated to CpG. (b) Representative immunofluorescence images of the lungs and mediastinal lymph nodes (MLNs) harvested from C57BL/6 mice (n = 3) at 48 hours after immunization with 20 μg of fluorescein amidite (FAM)-labeled (green) gp100 peptide or an equimolar amount of labeled amph-gp100. Scale bars, 100 μm. DAPI (blue) was used to stain nuclei. (c) Quantification of peptide signals from the lungs at 24 and 48 hours after the immunization in (b). (d) Quantification of peptide signals from the MLNs at 48 hours after the immunization in (b). (e) C57BL/6 mice (n = 5) were vaccinated with 5 μg of AL11 peptide combined with 8 μg of CpG or equimolar doses of amph-AL11 and amph-CpG administered intratracheally or subcutaneously on days 0 and 28 and then challenged with a lethal dose of SIV-Gag Vaccinia virus (106 PFU) intratracheally on day 150. (f) Animal weights over time after Vaccinia virus challenge in (e). (g) Animal survival after Vaccinia virus challenge in (e). (h) C57BL/6 mice (n = 10) were left naïve or vaccinated with 10 μg of gp100, 10 μg of Trp1, and 10 μg of Trp2 peptide antigens combined with 8 μg of CpG or vaccinated with equimolar doses of amph-peptides and amph-CpG on days 0 and 14. Thirty days after boost, mice were challenged with 4 × 105 B16F10 melanoma cells intravenously, and survival over time is plotted. In each cohort, the number of mice surviving at 80 days was indicated. Data are represented as means ± SEM from three independent experiments (e–g) and two independent experiments (b–d and h); *P < 0.05, **P < 0.001, and ****P < 0.0001 as determined by one-way ANOVA. Survival data were analyzed using a log-rank test. Reproduced with permission.13 Copyright 2021, American Association for the Advancement of Science.
