Panel A shows schematic diagram of sequential immunization with HIV 1 antigens over time without axes. Panel B shows line graph of serum anti TM4 core antibody levels with vertical axis anti TM4 core immunoglobulin E and horizontal axis days post prime. Panel C shows scatter plot of germinal center B cell percentage with vertical axis percentage of germinal center B cells of B220 positive cells and horizontal axis mouse genotypes. Panel D shows dot plot of combined variable heavy and variable light chain mutations with vertical axis number of mutations and horizontal axis mouse genotypes. Panel E shows scatter plot of clonal sequence proportions with vertical axis percentage of clonal sequences and horizontal axis mouse genotypes. Panel F shows bar graphs of immunoglobulin heavy and light chain variable gene usage with vertical axis frequency percentage and horizontal axis gene identities across mouse genotypes. Panel G shows dot plot of clone size distribution with vertical axis logarithmic clone size and horizontal axis mouse genotypes. Panel H shows scatter plot of inverse Simpson diversity index with vertical axis diversity index and horizontal axis mouse genotypes. Panel I shows scatter plot of Shannon entropy with vertical axis entropy score and horizontal axis mouse genotypes. Panel J shows scatter plot of TM4 binding germinal center B cells with vertical axis percentage of TM4 binding germinal center B cells and horizontal axis mouse genotypes.
Sequential immunization with an HIV-1 antigen series. (A) Diagram of the experimental protocol. (B) ELISA quantification of total anti-TM4 core serum antibodies measured weekly from days 6 to 69. Thin lines represent single mice; bold lines represent the group mean. (C) Percentage of GC B cells among B220+ B cells in the draining LN on day 96. Gating as in Fig. 2 E. Bars indicate the mean ± SD. (D) Dot plot showing combined VH + VL gene nucleotide mutations among GC B cell sequences on day 96. Each dot indicates an antibody VH + VL pair. Bars indicate the median. (E) Proportions of clonal sequences within each mouse at day 96. Each dot indicates sequences from one mouse. Bars indicate the median. (F) Bar graph showing the relative abundance of mouse Ighv (left panel) and Igkv (right panel) gene usage in GC B cells on day 96. Top 10 most frequent genes ranked by the mean frequency in mM-only mice are shown. Bars indicate the mean ± SD. (G) Dot plot depicting the percentage of clonal sequences from GC B cells on day 96. Each dot indicates a clone. Bars indicate the median. (H) Dot plot depicting inverse Simpson’s index of antibody sequences from day 96 GC B cells. Each dot represents one mouse. Bars indicate the median. (I) Dot plot showing Shannon’s entropy index from day 96 GC B cells. Each dot represents one mouse. Bars indicate the median. (J) Percentages of TM4-binding GC B cells. P values of nonparametric one-way ANOVA are indicated. ***P < 0.001.