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. 2008 Jun 9;205(6):1343–1355. doi: 10.1084/jem.20071572

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© 2008 Shimomura et al.

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Figure 8. — Requirement of BAFF-R–mediated signaling for intestinal CD21⁻CD23⁻ B cell development. (A) Expressions of BAFF signaling-related molecules in purified B cells from the large intestine of day 0 (open bars), 4 (gray bars), and 8 (black bars) after DSS treatment and from the spleen of day 0 (hatched bars) are shown. The results represent the means ± the SD from six individual mice in each group. *, P < 0.00 1 (comparison between colonic B cells at day 0 versus day 8). (B) Western blot analysis shows the expression levels of NF-κB2 p100 and p52 in purified B cells (>97% are IgM⁺) of the large intestine from 4–30 pooled WT mice at day 0 (D0, normal) and 8 (D8, recovery of DSS colitis), and of the spleen from day 0. The data are representative of two individual experiments. (C) Expressions of IgM versus CD21 or CD23 on the gated B220⁺ cells from normal large intestine of control and BAFF-R mutant mice are shown. (D–G) Intestinal inflammation was induced in BAFF-R mutant (A/WySnJ; Day 0, n = 5; Day 8, n = 16) and control (A/J) mice (Day 0, n = 5; Day 8, n = 10) by oral administration of 4% DSS for 4 d. (D and E) Proportion (CD3/IgM; D) and absolute numbers of IgM⁺ B cells (E) in the colon of control and BAFF-R mutant mice at day 8 of DSS colitis are shown. Presence of statistically significant (P < 0.001) compared with control mice at day 8 is indicated by **. (F and G) Large intestine from control mice (F) and BAFF-R mutant mice (G) at day 8 were subjected to immunohistochemical analysis for the detection of IgM⁺ cells. (H) Apoptosis of intestinal IgM⁺ B cells in control mice and BAFF-R mutant mice at day 8 of DSS colitis as judged by Annexin V staining is shown. The mean percentages of Annexin V⁺ cells among IgM⁺ cells are 36.6 ± 1.09% in control mice (n = 7) and 75.6 ± 1.64% in BAFF-R mutant mice (n = 7; P < 0.001). (I) Expression of AA4.1 versus B220 on the cells from normal intestine of control (left) and BAFF-R mutant (right) are shown. Bars, 100 μm.