Furthermore, Foxo1f/fCd19Cre mice had markedly fewer LN B cells a

Furthermore, Foxo1f/fCd19Cre mice had markedly fewer LN B cells and an increase in peripheral blood B cells (Supporting Information Fig. 1D). The paucity of LN B cells correlated with reduced surface expression of CD62L (L-selectin), the LN homing receptor (Supporting Information Fig. 1E). The mice also had a reduced percentage of CD5+ B cells in the peritoneal cavity (Supporting Information Fig. 1F). The report from Dengler et al. did not examine the developmental status or function of peripheral B220+IgM+ cells in Foxo1f/fCd19Cre mice 10. We stained splenocytes from our Foxo1f/fCd19Cre mice and

controls with antibody combinations that distinguish two mature subsets (FO, MZ) and four transitional Anti-infection Compound Library concentration B-cell subsets (T1, T2, T3 and MZ precursor (MZP)) 13. When compared with control Foxo1f/+Cd19Cre mice, Foxo1f/fCd19Cre mice displayed a consistent and statistically significant increase in the percentage of MZ cells, defined as B220+AA4.1−IgMhiCD21hiCD23lo (Fig. 1A). In contrast, the percentage of FO cells (B220+AA4.1−IgMloCD21intCD23hi) was reduced (Fig. 1A). A normal percentage of MZP cells was present in Foxo1f/fCd19Cre mice, despite reduced percentages of T1 and T2 cells; this suggests that immature transitional cells might commit preferentially to the MZP stage. The absolute numbers of splenocytes were equivalent between Foxo1f/fCd19Cre mice and control mice (data not shown). Increased abundance of B220+ cells

in the splenic MZ and other extrafollicular regions https://www.selleckchem.com/products/ulixertinib-bvd-523-vrt752271.html was also apparent by immunofluorescent staining of spleen sections (Fig. 1B). The

percentages of mature FO and MZ cells were comparable in the two control groups (Foxo1f/+Cd19Cre and Foxo1f/f) (Fig. 1A), and other experiments showed a consistently greater population of MZ cells (B220+CD21hiCD23lo) in Foxo1f/fCd19Cre compared with Foxo1f/f mice (data not shown). Therefore, we used Foxo1f/f mice as controls in Fig. 1B and in other experiments to simplify breeding schemes. The altered balance of FO and MZ cells in Foxo1f/fCd19Cre mice Carbohydrate was not observed in analyses of mice with Foxo1-deficient B cells generated using Cd21Cre10. A likely explanation is that Cd21Cre drives deletion of Foxo1 at a time point after transitional B cells commit to either the FO or the MZ lineage, whereas Cd19Cre deletion is complete by this stage. Interestingly, Foxo1f/fCd21Cre mice 10 shared the reduced LN B-cell population and CD62L expression observed here in Foxo1f/fCd19Cre mice. This could be explained by a requirement for Foxo1 in CD62L gene expression in mature B cells, after Cd21Cre-mediated deletion is completed. We purified splenic B cells and activated them in vitro with titrated doses of either a BCR stimulus (anti-IgM) or a TLR stimulus (LPS). We measured cell proliferation and survival by cell division tracking using CFSE. B cells from Foxo1f/fCd19Cre proliferated more weakly to anti-IgM, compared with B cells from Foxo1f/f mice (Fig. 2A).

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