Fig 2: Effect of pre-phase B cell depletion on innate and adaptive immune cell subsets in autoimmune BXSB mice(A and B) Representative FACS plots (A) and the quantification (B) of immature (Imm. B), follicular (FO), and marginal zone (MZB) B cell subsets in the spleen of 18-week-old female (?) or male (?) BXSB mice treated with isotype control or CD20-specific antibodies during the pre-phase of disease. Shown are two independent experiments with 5–7 mice per group. Differences between groups were determined by ordinary one-way ANOVA with Tukey's multiple comparison test. Symbols represent individual mice; error bars represent the mean ± SD. **p < 0.01; ***p < 0.001.(C–F) Depicted are representative FACS plots (C, E) and the quantification (D, F) of CD138+ TACI+ plasma cells in the spleen (C, D) and bone marrow (E, F) of 18-week-old female (?) and male (?) BXSB mice treated with isotype control or CD20-specific antibodies during the pre-phase of disease. Shown are two independent experiments with 5–7 mice per group. Differences between groups were determined by ordinary one-way ANOVA with Tukey's multiple comparison test. Symbols represent individual mice; error bars represent the mean ± SD. **p < 0.01; ***p < 0.001; ****p < 0.0001.(G–J) Shown are representative FACS plots (G, I) and the quantification (H, J) of CD4+ (G, H) and CD8+ (I, J) T cell subsets (naive, effector memory (EM), central memory (CM)) in the spleen of 18-week-old female (?) and male (?) BXSB mice treated with isotype control or CD20-specific antibodies during the pre-phase of disease. Shown are two independent experiments with 5–7 mice per group. Differences between groups were determined by ordinary one-way ANOVA with Tukey's multiple comparison test. Symbols represent individual mice; error bars represent the mean ± SD. *p < 0.05; ***p < 0.001.(K and L) Depicted are representative FACS plots (K) and the quantification (L) of classical and non-classical monocyte subsets in the spleen of 18-week-old female (?) and male (?) BXSB mice treated with isotype control or CD20-specific antibodies during the pre-phase of disease. Shown are two independent experiments with 5–7 mice per group. Differences between groups were determined by ordinary one-way ANOVA with Tukey's multiple comparison test. Symbols represent individual mice; error bars represent the mean ± SD. **p < 0.01; ****p < 0.0001.

Fig 3: Effect of pre-phase B cell depletion on lymphadenopathy and splenomegaly(A and B) Depicted are representative pictures of inguinal, axial, and cervical lymph nodes (A) and spleens (B) from isotype or anti-CD20 antibody (aCD20) treated male (?) and female (?) mice at 18 weeks of age of two independent experiments.(C) Quantification of the spleen weight of 18-week-old anti-CD20 or isotype control antibody-treated BXSB mice. Significant differences between indicated groups were tested with ordinary one-way ANOVA with Tukey's multiple comparison test. Shown are the results of two independent experiments with a total of n = 5–6 mice per group. Symbols represent individual mice; error bars represent the mean ± SD. ****p < 0.0001.(D and E) Shown are representative immunofluorescence pictures of lymph node (D) and spleen (E) sections from 18-week-old female (?) and male (?) BXSB mice stained for the presence of B220+ B cells (depicted in green), TCRß+ T cells (in blue) and CD138+ plasma blasts and plasma cells (in red). Size bars in D indicate 200 µm (2000 µm for isotype-treated male BXSB mice). Size bars in E indicate 500 µm.