Fig. 5: Expansion and diversification of polyreactive clones.
a, Clones with descendants in total repertoire. Graph shows the number of the in vitro expressed clones by donors (gray bars). Number of clones with identified descendants in total repertoire are depicted with deep red. Data are shown from HC-Ags (n = 3), pRD-N (n = 1) and patients with pRD-Ag (n = 5) with individual IDs indicated. b, Contribution of descendants to repertoire. Fraction of the descendants of clones in total repertoires are shown by individuals. Descendants of the polyreactive and non-polyreactive clones are depicted with filled and striped bars, respectively. c, Compartmental distribution of the descendants. The contribution of the descendants of each clone to the total repertoires are shown in the indicated individuals by CD38int (blue); CD38− (red) and CD27+ (green) compartments. Clone IDs are shown; deep red and black represents polyreactive and non-polyreactive clones, respectively. Striped bars for P14 represent clones identified in repertoires obtained 1 year later after isolation of original clones. Clones with shared descendants in the two time points are indicated with bold. d, Single nucleotide mutations in clones. Average number of mutations in the V segment by individuals are shown as mean ± s.e.m. for HCs (n = 3), pRD-Ag (n = 5) and pRD-N (n = 1) with individual values depicted. Statistical analyses were performed on individuals with antigens (HC-Ag versus pRD-Ag) using a Mann–Whitney U-test. e, Phylogenic lineage trees of expanded clonotypes. Immunoglobulin trees represent expansion of individual clonotypes by their SHM profile. Putative unmutated common ancestors (UCAs) are depicted by squares, arrowhead indicates the original in vitro expressed sequence, descendants are shown by CD38int (blue), CD38− (red) or CD27+ (green) compartments and the color of the nodes indicates antigen selection strength (blue to deep red). Size of the nodes represent sequence abundance. The number of the nucleotide difference between adjacent sequences and the isotype of each sequence are shown. Lineages derived from polyreactive or non-polyreactive CD38int clones are depicted with deep red or black, respectively. Orange line indicates descendants from second time point for P14.
