Lineage tracing of pt7 bNAbs in the blood and bone marrow. (A) Plots show divergence/identity analysis of pt7 IgG and IgA heavy chain (IgH) sequences obtained by Ig-HTS from peripheral blood (B) and bone marrow (BM) mononuclear cells. IgH sequences filtered on bNAb-specific V-J rearrangements (total numbers are shown below the plots) are depicted as function of the identity to bNAb reference sequence (% bNAb-id) and of divergence to inferred GL sequence (% GL-div). Gate numbers indicate the % of bNAb-relative sequences. (B) Divergence/identity plots showing the overlay of 7-176 and 7-269 sequences from pt7 blood and bone marrow IgA and IgG DNA libraries as shown in A. (C) Phylogenic tree (right) shows the relationship between 7-269 clonally related IgH nucleotide sequences found by high-throughput sequencing in blood and bone marrow IgA-expressing cells (in dark and light blue, respectively) and the bone marrow IgG repertoire (in light red) as shown in B, and by single B cell gp160-capture (as shown in Fig. 1 D, blue) used as reference sequences (in black). Only bone marrow 7-269–related sequences with GL divergence ≤4% were included in the phylogenic analysis. (D) Sequence logo showing the alignment of the CDR_H3s from the V_H sequences obtained by Ig-HTS shown in C (n = 131). The frequency of Cys residues in position 105 and 110 is shown below.