Figure 3. Vaccine nanodiscs for LN-targeting of Ag and adjuvants and elicitation of CTL responses.
a-b, C57BL/6 mice were administered subcutaneously at tail base with a, 31 nmol FITC-tagged Ag (CSSSIINFEK(FITC)L) or b, 2.3 nmol Cho-CpG (20% labeled by Cy5) in free soluble or sHDL form, and fluorescence signal in the draining inguinal LNs were quantified with IVIS after 24 h. c-e, C57BL/6 mice were immunized with the indicated formulations (15.5 nmol Ag peptide and 2.3 nmol CpG) on days 0, 21, and 42. c, Shown are their representative scatter plots on day 49 and d the frequency of SIINFEKL-specific CD8α+ T-cells in peripheral blood measured 7 days post each immunization by flow-cytometry analysis of tetramer+ CD8α+ T-cells. e, On day 50, pre-vaccinated animals were challenged with subcutaneous flank injection of 2×105 B16OVA cells, and tumor growth was measured over time. f-h, C57BL/6 mice were immunized in a biweekly interval. Shown are f, percent of SIINFEKL-specific CD8α+ T-cells in peripheral blood; g, ELISPOT analysis of IFN-γ spot-forming cells among splenocytes after ex vivo restimulation with SIINFEKL on day 35; and h, Ag-specific CD8α+ T-cell responses measured over 12 weeks post vaccination (black arrows indicate days of immunizations). i, Vaccinated mice in (h) were intravenously challenged with 5×104 B16OVA cells two months after the third vaccination. Shown are pictures of the lungs and numbers of lung metastatic nodules counted on day 20 after the B16OVA challenge. The data show mean ± SD from a representative experiment (n = 4-5) from 2-3 independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001, analyzed by (a-b) two-tailed unpaired Student’s t test, (d,e,f,h) two-way ANOVA, or (i) one-way ANOVA with Bonferroni multiple comparisons post-test. Asterisks in panel e indicate statistically significant differences between sHDL-Ag/CpG and SIINFEKL+CpG+Montanide.