Figure 3.

Diversity of Ab responses to TT and Plasmodium Ags. B cells specific for TT (n = 27 cells from 2 donors), PfCSP (n = 31 cells from three donors) and PfMSP1 (n = 31 cells from three donors, one sort per donor) were sorted after tetramer staining and rearranged Ig V(D)J sequences and constant regions determined by RNA-seq. Analysis of (A) Ig isotype and (B) IGHV gene use by Ag-specific B cells; note for some cells it was not possible to determine the isotype used. (C) Analysis of clonal relationships between B cells specific for each Ag; each wedge constitutes a unique clone, clones with multiple representatives are coloured and if a clone spans two phenotypes the link is indicated. (D) Analysis of mutation frequency by cell phenotype; bars show mean ± SEM; analysis was by one-way ANOVA with Tukey post-test controlling for subject as a random effect. (E) Association of mutation frequency with Ag and Ab isotype; analysis by two-way ANOVA controlling for subject as a random factor with Tukey post-test, bars show mean ± SEM details of the model are given below the graph, significant pairwise comparisons indicated. (F) Results of BASELINe analysis of selective pressure (sigma) on Ig gene sequences in (i) CDRs and (ii) FWRs; mean ± SEM shown for each Ag, analysis by single sample ANOVA to determine if the selective pressure is significantly different from zero.