Fig. 4: 10E8 B cells do not persist in GCs.

a, Flow cytometry plots of CD45.2⁺ GC B and CD45.1⁺ GC B cells at days 7, 14 and 21 p.i. in the spleens of wild-type CD45.1 mice adoptively transferred (intravenously) with 200,000 CD45.2 10E8-UCA^H B cells to establish a frequency of 1:10⁴ 10E8-UCA^H B precursors and immunized 1 day later with 5 µg of 10E8-GT10.2 12mer (n = 3 mice) or 5 µg of 10E8-GT9-KO 12mer (n = 3) with alhydrogel. Representative data from one of three independent experiments are shown. b, Percentage of CD45.2⁺ GC B cells in splenic GCs (top) and percentage of CD45.1⁺GT10⁺⁺KO⁻ GC B cells in splenic GCs (bottom) at days 7, 14 and 21 p.i. as in a (n = 3). Each circle represents one mouse. Top, representative data from one of three independent experiments; lines mark the means. Bottom, data are from pooled groups from three independent experiments; lines mark the respective means. c, Amino acid SHM of 10E8-UCA^H CD45.2⁺GT10⁺⁺KO⁻ B cells isolated from spleen GCs at days 7 and 14 p.i. with 10E8-GT10.2 12mer, endogenous CD45.1⁺GT10⁺⁺KO⁻ B cells isolated from spleen GCs at day 14 p.i. with 10E8-GT10.2 60mer and pretransfer (day −1, naive) 10E8-UCA^H GT10⁺⁺KO⁻ and wild-type GT10⁺⁺KO⁻ B cells as a control. Circles represent individual sequences, and lines indicate median values. d, Affinities of monomeric mAbs expressed from 10E8-UCA^H CD45.2⁺GT10⁺⁺KO⁻ B cells (GT10.2-10E8-UCA^H), endogenous CD45.1⁺GT10⁺⁺KO⁻ B cells (GT10.2-WT) and pretransfer (day −1) 10E8-UCA^H GT10⁺⁺KO⁻ B cells and wild-type GT10⁺⁺KO⁻ B cells (naive), as in c, determined by monomer SPR assay. Circles represent individual antibodies, and lines represent median values; LOD, limit of detection.