Fig. 4: Plasma cells, B.63521 gp120-specific LLPCs, and memory B cells in the bone marrow.

A The frequency of CD138⁺⁺CD38⁺⁺CD20^-CD19⁺ plasma cells at weeks -4, 26, and 56 are presented as mean ± SEM. Each individual NHP is represented as a symbol (N = 7/group). B The percentage of gp120 protein-specific plasma cells in the bone marrows collected at week 56, positive for intracellular dual-fluorophore labeled B.63521 gp120 proteins (gp120-BV421 and gp120-PE). The right panels show the representative flow plots. The flow panel and the gating strategy are shown in Table S8 and Fig. S5A, respectively. C Spearman correlation between B.65321 gp120-specific bone marrow plasma cell frequency and antibody avidity score at week 56 is shown. D Memory B cell ELISpot images of duplicate or triplicate wells from bone marrow cells from one or two animals in the group, depicted at week 48 following in vitro stimulation of BM cells. The B.63521 gp120 protein-specific IgG memory B cell ELISpot images following stimulation are shown for naïve, AH, and ALFQ groups. E The number of memory B cell B.63521 gp120-specific IgG and F IgA spot-forming units/10⁶ bone marrow cells are presented as mean ± SEM. Each symbol represents the average of triplicate or duplicate wells for each NHP from each group. The responses were considered positive when the spot count exceeded the mean ± 3 SD of the negative control wells. Statistical comparisons between the different adjuvant groups were performed using the Kruskal–Wallis test with Dunn’s multiple comparison test. *p < 0.01, **p < 0.008, and ***p < 0.0002.