Figure 3.

Only HIV-1 Env is subdominant to the nanoparticle scaffold in a series of different nanoparticle immunogens that all use the same I53-50 scaffold

(A) Schematic representation of the series of nanoparticle immunogens used in this study that all use the same I53-50 scaffold, highlighting the structural differences in the displayed antigen for each immunogen (antigen in blue, glycans in green, I53-50A trimeric component in gray, I53-50B pentameric component in orange).

(B) Table listing the nanoparticle and non-assembling control immunogens and schematic depicting the study timeline and blood collection time points that each data panel represents.

(C and D) Antigen-specific (C) and I53-50 scaffold-specific (D) serum IgG binding titers in BALB/c mice immunized with the color-coded immunogens listed in the table in (B), measured by ELISA and plotted as the area under the curve (AUC) for each serum dilution series. Antigen-specific IgG titers were measured by Ni-NTA-capture ELISA for more accurate comparison among immunogen groups. Each symbol represents an individual animal, and the geometric mean AUC from each group is indicated by the bar (N = 10 mice/group). The blue dashed line in (D) represents levels for the ConM-I53-50 immunogen for comparison.

(E) Ratio of the antigen-specific (C) to I53-50 scaffold-specific (D) binding antibody AUC titers. The black dashed line indicates a ratio of 1.

(F) Spearman’s correlations between post-second boost (week 10) anti-antigen and anti-I53-50 scaffold serum IgG titers (AUC) for all immunogens on the same plot. Shaded areas represent 95% confidence intervals of the plotted linear regression line. Each symbol represents a mouse (N = 10 per immunogen).

p values between groups were determined by Brown-Forsythe and Welch one-way ANOVA test, with Dunnett’s T3 multiple comparisons test. ns, non-significant; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001.