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. 2023 Nov 3;211(12):1814–1822. doi: 10.4049/jimmunol.2300607

FIGURE 5.

FIGURE 5.

The CD40 TRAF2 binding motif but not the TRAF6 binding motif is required for TD GC responses. WT and CD40 mutant mice were immunized with NP-KLH in alum. After 8 d, GC B cells and total B cells were analyzed in the spleen. Representative FACS analysis (A) of CD38−GL7+ GC B cells in the CD4− B220+ B cell gate and statistical analyses of FACS data examining the total number of spleen B cells and GC B cells in each mouse strain (B). (C) OT-II T cells (CD45.1) were transferred to the indicated recipient mice (CD45.2) and were immunized with NP-OVA/alum 1 d after cell injection. Transferred OT-II cells were analyzed at day 8 by FACS analysis. A representative flow cytometry profile of spleen OT-II cells (CD45.1+CD4+B220−) analyzed for the Tfh CXCR5high PD-1high phenotype and statistical analyses of FACS data showing the percentage of transferred OT-II cells expressing a Tfh phenotype are shown. (D) WT and CD40 mutant mice were immunized with NP-KLH in alum. Serum was collected 3 wk after immunization, and total NP-specific Ab (anti-NP30 IgG) and high-affinity NP Ab (anti-NP2 IgG) was determined by ELISA. Data are combined from two independent experiments (mean ± SD using six mice per group). One-way ANOVA followed by Dunnett test against the immunized WT group was performed for multiple comparisons. **p < 0.01, ***p < 0.001, ****p < 0.0001.