Extended Data Fig. 10: Mice carrying native human precursor sequence show a sustained GC response and form memory, related to Fig. 7.
From: Affinity gaps among B cells in germinal centers drive the selection of MPER precursors

a, Schematic of CD45.1+ mice adoptively transferred to establish a frequency of 1:104 CD45.2+MPER-HuGL18H B cells and then immunized with either 5 μg of 10E8-GT10.3 12mer immunogen and alhydrogel, or 10E8-GT9-KO 12mer (control), as per Fig. 7b. b, Quantification of epitope specific CD45.2+MPER-HuGL18H B cells in mice immunized with 10E8-GT10.3 12mer as in a. Error bars indicate mean ± SD from mice in each group (n = 6–8). Data pooled from two independent experiments. c, Immunohistochemistry of spleen at 21 DPI. Red: GL7 (GC marker); White: CD45.2; Green: IgD; Blue: TCRβ. Scale, 250 um. d, Quantification showing CD45.2 recruitment in GCs from 10E8-UCAH or MPER-HuGL18H transferred mice immunized with 10E8-GT10.2 12mer and 10E8-GT10.3 12mer (merged data from Figs. 6d, 7c, and ref. 25). Data are pooled from 2–3 independent experiments (n = 4–6). Error bars indicate mean ± SD. (*p = 0.0131, **p = 0.0085) (unpaired t test, two-tailed). e, Representative flow cytometry showing gating strategy used for the identification of memory B cells (MBC) 36 DPI with 10E8-GT10.3 12mer with alhydrogel (n = 8) or 10E8-GT9-KO 12mer with alhydrogel (n = 5). f, Quantification of memory B cells (MBC) 36 DPI with 10E8-GT10.3 12mer, as in e. Experiment performed once. Error bars indicate mean ± SD. (p = 0.024) (unpaired t-test, two-tailed). g, Graph of the affinities of MPER-HuGL18H mAbs 21 DPI with 10E8-GT10.3 12mer (panel with affinities) (y-axis) sorted by light chain (x-axis) LOD: limit of detection. h, Graph showing the affinities (y-axis) of the subset using IGKV1-117 light chains from the MPER-HuGL18H 21 DPI with 10E8-GT10.3 12mer (selection from Fig. 7g) vs. number of heavy chain mutations (x-axis). LOD: limit of detection. i, Tree showing phylogenetic relationship between MPER-HuGL18H B cell IGHV sequences, colorized on the basis of 21 DPI affinities shown in (Fig. 7g).