Fig. 6
Primary and memory T cell-dependent responses of single-κ and dual-κ B cells. a Representative analysis of PE-binding single and dual-κ cells within CD22+Igκ+ splenocytes (gated as in Supplementary Fig. 8b) of MRL/lpr-Igkm/h mice 7 days after injection of PBS, PE + Alum, or PE + CpG. b Frequencies of PE-binding single and dual-κ cells (analyzed as in panel a) from MRL/lpr-Igkm/h mice, 7 days after injection of PBS or PE + Alum. Data represent fold changes relative to PE-reactive single-κ B cells in PBS-injected mice. c Ratio of dual-κ to single-κ PE-binding cells in MRL/lpr-Igkm/h mice treated as indicated. Data in (b, c) are combined from four independent experiments with 6–7-wk-old MRL/lpr-Igkm/h mice injected with either PBS (N = 9) or PE + Alum (N = 11). d Frequencies of PE-binding single and dual-κ cells (analyzed as in panel a) from wt (N = 6) and IL-21R KO MRL/lpr-Igkm/h (N = 5) mice 7 days after injection of PE + Alum, shown as fold change relative to PE-reactive single-κ B cells in naive mice of corresponding strain (N = 3 and N = 6, respectively, combined from two experiments). e Frequencies of PE-binding single or dual-κ B cells (analyzed as in panel a) from MRL/lpr-Igkm/h mice immunized with either PE + Alum or PE + CpG, shown as fold change relative to single-κ B cells in PBS-injected mice. f Ratio of dual-κ to single-κ B cells in mice immunized as indicated. Data in (e, f) are combined from two independent experiments using 6–12-wk-old MRL/lpr-Igkm/h mice immunized with PE + Alum (N = 4 total) or PE + CpG (N = 6 total). g Schematics of experiment to measure memory response of PE-binding B cells. h Frequencies of PE-binding single or dual-κ B cells (analyzed as in panel a) from control MRL/lpr-Igkm/h mice receiving only a primary (PE + Alum, N = 3) or secondary (PE, N = 7) immunization, and experimental mice receiving both primary and secondary immunizations (N = 6), combined from three independent experiments. Single and dual-κ cells for control mice are shown together. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; n.s. not significant. Significance was assessed by Student’s t- or Mann–Whitney tests. In all bar graphs the standard error bars represent SEM and each symbol represents an individual mouse
