Fig. 4

Serum IgG specificity, neutralizing activity, and effector function. Binding of IgG antibodies at two weeks post the first cycP-gp120 immunization (wk 25) to a gp120 and gp140 antigens and b gp70-V1V2 scaffolds representing the global diversity of HIV-1 determined using Binding Antibody Multiplex Assay (BAMA). Shaded areas indicate clade. c Binding of peak immune sera (wk 25) to 15mer peptides derived from clade B consensus gp120, measured by peptide microarray linear epitope mapping and reported as binding signal (Log2 fold difference post-immunization/baseline binding intensity). Magnitude of binding to each epitope is defined as the highest binding signal for a single peptide within the region of the epitope. d Representation of peptide array binding of each clade B consensus epitope as % of the total response. e Anti-V2 hotspot (HS) response (wk 25) against V2-HS peptides derived from strains JRFL (E168K), ADA, and SF162P3 measured by ELISA. Dotted lines connect data from same animal. Blue squares, needle-free SL/B; gray triangles, ID/SC. f Neutralizing antibodies over time against HIV-1 isolates MW965.26, SF162.LS, and BaL.26, measured as ID50. g Antibody-dependent phagocytosis (ADP) at pre-challenge (wk 45). ADP score (mean ± S.D.) calculated for each serum by dividing the median fluorescence intensity (MFI) of bead positive cells by the value obtained using the same dilution of pooled serum from naive macaques. h Antibody-dependent cell viral inhibition (ADCVI) measured at pre-challenge as % viral inhibition (mean ± S.D.). An average of two individual experiments is shown. Box and whiskers plots (a–c, f); box extends from 25th to 75th percentile, line indicates median, whiskers indicate min and max values (*p < 0.05; **p < 0.01; ***p < 0.001; ****, p < 0.0001; 2-Way ANOVA, multiple comparisons). ID50, serum dilution required to neutralize 50% infection