Figure 6.
Analysis of splenic GC, IgG1 class switched B cells, and PC differentiation after the fifth immunization boost. (A) Schematic representation of the induction and immunization protocol for the mid-term cohort. (B) Strategy for gating TOM+ B cells from TOM-AID mice (upper panel) and TOM+/YFP− and TOM+/YFP+ from REL-AID mice (lower panel) as well as GCBs from total CD19+ live B cells (CD19+ B220+/−). (C) Percentage of GCBs among TOM+ B cells in AID-TOM control and REL-AID mice. Mean and SEM are shown by a red and 2 black lines, respectively. The P value of the Mann-Whitney test is shown. (D) Percentage of TOM+/YFP+ B cells in the total TOM+ (left) or in the GCB (right) fraction in REL-AID mice. Each line connects the points for 1 mouse. The P value of the paired Wilcoxon test is shown. (E) Example of gating on IgM−/IgD− double-negative (DN) B cells in both AID-TOM (upper) and REL-AID (lower) mice. (F) Analysis of DN B cells in both AID-TOM and REL-AID mice. Upper: ratio of DN/IgM+ cells among total TOM+ B cells in AID-TOM and REL-AID mice. Lower: percentages of YFP+ B cells among TOM+ B cells in the total TOM+ and in the DN B-cell fraction in REL-AID mice (each line connects the points for 1 mouse). The P values of the Mann-Whitney (upper) and paired Wilcoxon test (lower) are shown. (G) Example IgG1 class switched B-cells gating for AID-TOM (upper) and REL-AID (lower) mice. (H) Analysis of IgG1+ B cells in both AID-TOM and REL-AID mice. Left: percentages of IgG1+ (also IgM− and IgD−) B cells among TOM+ B cells in AID-TOM and REL-AID mice. Right: percentages of YFP+ B cells among total TOM+ and IgG1+ B cells (each line connects the points for 1 mouse). The P values of the Mann-Whitney (upper) and paired Wilcoxon test (lower) are shown. Each experiment has been done at least 4 times.

Analysis of splenic GC, IgG1 class switched B cells, and PC differentiation after the fifth immunization boost. (A) Schematic representation of the induction and immunization protocol for the mid-term cohort. (B) Strategy for gating TOM+ B cells from TOM-AID mice (upper panel) and TOM+/YFP− and TOM+/YFP+ from REL-AID mice (lower panel) as well as GCBs from total CD19+ live B cells (CD19+ B220+/−). (C) Percentage of GCBs among TOM+ B cells in AID-TOM control and REL-AID mice. Mean and SEM are shown by a red and 2 black lines, respectively. The P value of the Mann-Whitney test is shown. (D) Percentage of TOM+/YFP+ B cells in the total TOM+ (left) or in the GCB (right) fraction in REL-AID mice. Each line connects the points for 1 mouse. The P value of the paired Wilcoxon test is shown. (E) Example of gating on IgM−/IgD− double-negative (DN) B cells in both AID-TOM (upper) and REL-AID (lower) mice. (F) Analysis of DN B cells in both AID-TOM and REL-AID mice. Upper: ratio of DN/IgM+ cells among total TOM+ B cells in AID-TOM and REL-AID mice. Lower: percentages of YFP+ B cells among TOM+ B cells in the total TOM+ and in the DN B-cell fraction in REL-AID mice (each line connects the points for 1 mouse). The P values of the Mann-Whitney (upper) and paired Wilcoxon test (lower) are shown. (G) Example IgG1 class switched B-cells gating for AID-TOM (upper) and REL-AID (lower) mice. (H) Analysis of IgG1+ B cells in both AID-TOM and REL-AID mice. Left: percentages of IgG1+ (also IgM− and IgD−) B cells among TOM+ B cells in AID-TOM and REL-AID mice. Right: percentages of YFP+ B cells among total TOM+ and IgG1+ B cells (each line connects the points for 1 mouse). The P values of the Mann-Whitney (upper) and paired Wilcoxon test (lower) are shown. Each experiment has been done at least 4 times.

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