|Figure 5: Absence of PD-L1 but not PD-L2 leads to a significant decrease in Breg cells.
Splenocytes from control and E2-implanted WT, PD-L1-/- and PD-L2-/- mice were harvested on day 24 post-immunization and assessed for expression of: A) CD1dhiCD5+ B cells, C) PD-L1-expressing B cells and D) PD-L2-expressing B cells Data represent CD1dhiCD5+, PD-L1+ and PD-L2+ expression on gated CD19+ cells of total gated and live cells. Data are representative of 2 independent experiments with spleens processed from 6-8 individual mice (mean ± SEM). B) Splenocytes from control and E2-implanted WT, PD-L1-/- and PD-L2-/- mice, harvested on Day 24 post-immunization, were cultured in the presence of 25 μg/ml mMOG35-55 peptide for 48 h. Cells were pooled from mouse groups (6-8 mice total; 2 individual experiments) (mean ± SEM) and cytokines were detected using the Luminex bead array. Significant differences between the groups (*p ≤ 0.05, **p ≤ 0.01 and ***p ≤ 0.001) were determined using One-way ANOVA with one way analysis of variance and Newman-Keuls Multiple Comparison post test and are indicated by brackets.