Fig. 7

Plasma cell differentiation is enhanced by transgenic CD30 expression. A FACS plots illustrating the gating strategy for activated FoB cells (CD43⁺CD23⁺) and plasmablasts + B1 cells (CD43⁺CD23⁻). The FACS plots were pregated as shown in Supplementary Fig. 3a. CD19⁺ cells were subsequently gated on reporter⁺ cells. The graphs summarize the percentages of reporter⁺ activated FoB cells and CD43⁺CD23⁻ B cells in the spleen at the indicated time points after immunization. CD30//Cγ1-Cre mice (red dots), control mice (CAR//Cγ1-Cre mice; blue triangles) (N = 3–13 per group). Rounds of immunization: day 4: 5; day 7: 4; day 11: 4; day 21: 2. B Gating strategy for plasmablasts (IRF4⁺B220^low) in the spleen. The FACS plots were pregated as indicated in Supplementary Fig. 5b. The cells in the large lymphocyte gate were subsequently gated on reporter⁺ cells. The graph compiles the percentages of IRF4⁺B220^low cells in CD30//Cγ1-Cre mice (red dots) and controls (CAR//Cγ1-Cre mice; blue triangles) on day 7 and day 11 after immunization (N = 3–11 per group). Rounds of immunization: day 7: 2; day 11: 7. C Immunohistochemistry images depict the localization of IRF4⁺ cells (red) in histological sections from the spleen on day 11 after immunization. B cells were stained with B220 (green), and the marginal sinus was stained with anti-Laminin (blue). D FACS plots of the gating strategy for plasma cells (TACI⁺B220^low) are shown. The FACS plots were pregated as indicated in Supplementary Fig. 5b. The cells in the large lymphocyte gate were subsequently gated on reporter⁺ cells. The graphs summarize the percentages of reporter⁺ plasma cells at days 7 and 11 after immunization (N = 6–11 per group). Rounds of immunization: day 7: 4; day 11: 6. E The graphs compile the percentages of reporter⁺ B cells (left), reporter⁺ plasmablasts and B1 cells (CD43⁺CD23⁻) (middle), and reporter⁺ plasma cells (CD138⁺B220^low) (right) in the blood. CD30//Cγ1-Cre mice (red dots) and controls (CAR//Cγ1-Cre mice; blue triangles) are shown (N = 9–11 per group). Rounds of immunization: 5. F The serum titers of NP-specific antibodies were determined by ELISA using NP3-BSA (for high-affinity NP-specific antibodies) or NP13-BSA (for total NP-specific antibodies) as the capture reagent. The ratios of high (binding to NP3) to total NP-specific (binding to NP13) antibodies for the IgM and IgG1 subclasses were determined on day 7 and day 11 after immunization (N = 4–8 per group). In the last row, the ratios of NP3/NP17 from day 7 and day 11 were combined. The relative titers of anti-NP-IgM and anti-NP-IgG1 antibodies bound to NP-13-BSA and NP-3-BSA in the sera of CD30//Cγ1-Cre mice and control mice are shown in Supplementary Fig. 9b. 2-way ANOVA with Sidak’s (A) or Tukey’s (D) multiple comparisons test. B + F, Unpaired t tests. E Unpaired t test. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001