Fig. 3: Antigen formulation significantly enhances epitope-specific 10E8 B cells in GCs.

a, Affinities of 10E8-UCA^H/mouse^L Fabs or human 10E8-UCA^H/UCA^L for 10E8-GT9.2, 10E8-GT9.6 and 10E8-GT10.2 12mers in nanoparticle SPR assays. Symbols represent individual antibodies, and lines represent 10E8-UCA^H/mouse^L median values. b,c, Representative flow cytometry plots of CD45.2⁺ and CD45.1⁺ GC B cells (b) and percentage of GC B cells and CD45.2⁺ GC B cells (c) at day 7 p.i. in the spleens of CD45.1 wild-type mice transferred with CD45.2 10E8-UCA^H B cells (frequency of 1:10⁴ CD45.2⁺ 10E8-UCA^H B cells) and immunized 1 day later with 50 µg of 10E8-GT9.6 12mer, 10E8-GT10.2 12mer and 10E8-GT9-KO 12mer with Ribi adjuvant. Error bars indicate mean ± s.d. from mice in each group (n = 4–6). Significance was calculated with a two-tailed, paired t-test. Data shown are from one of two independent experiments; NS, not significant. d, Representative flow cytometry plots (left) and quantifications (right) of 10E8-UCA^H CD45.2⁺ B cells in GC B cells at day 7 p.i. in the spleens of CD45.1 wild-type mice transferred with CD45.2 10E8-UCA^H B cells (frequency of 1:10⁴ CD45.2⁺10E8-UCA^H B cells) immunized with 10E8-GT10.2 12mer (40 µg, 10 µg and 2.5 µg) or 10 µg of 10E8-GT9-KO 12mer with Ribi adjuvant (n = 4; top) or with 10E8-GT10.2 12mer (20 µg, 5 µg and 1.25 µg) or 5 µg of 10E8-GT9-KO 12mer with alhydrogel adjuvant (bottom). Error bars indicate mean ± s.d. from mice in each group (n = 3). Alhydrogel data are representative of three independent experiments, and Ribi data are representative of two independent experiments, with one quantified. e, Representative immunohistochemistry image of the spleen (top) and percentage (bottom) of 10E8-UCA^H CD45.2⁺ B cells in GCs in recipients at day 7 p.i., as in d. Scale bar, 50 µm. The image is representative of images obtained from four mice.