Figure 6

Serum anti-DNA antibody levels, DNA-reactive B cells and IFNγ transcripts in WT, ERα-deficient and ERβ-deficient mice. (A) Serum anti-DNA antibody levels in WT, ERα-deficient and ERβ-deficient mice. E2 or placebo (P) was administered to R4A-Tg WT, ERα-deficient and ERβ-deficient mice for 6 wk. Serum was obtained at several time points and analyzed for anti-dsDNA antibody levels. E2 induced increased anti-DNA antibody titers in WT and ERβ-deficient mice, whereas placebo treatment led to no change in antibody titer. ERα-deficient mice treated with E2 failed to display an increase in anti-DNA antibody titers compared with placebo. (B) Enumeration of DNA-reactive B cells in WT, ERα-deficient and ERβ-deficient mice. E2 or placebo was administered to WT, ERα-deficient and ERβ-deficient mice for 5–6 wk, and the total number of splenic B cells producing γ2b and B cells producing γ2b anti-dsDNA antibody was quantitated by ELISpot assay. The frequency of anti-dsDNA B cells among the γ2b-producing B cells was calculated as DNA spots/γ2b spots. Both WT and ERβ-deficient mice displayed an increased frequency of DNA-reactive B cells after E2 administration compared with placebo administration; ERα-deficient mice showed no E2-induced change in DNA-reactive B cells. Five mice were used per group for these studies. (C) Type 1 (IFNα, IFNβ) and type 2 (IFNγ) transcripts were measured in total splenocytes from ERα^-/-, ERβ^-/- and WT mice. The relative expression in comparison to Polr2A is represented. Six or eight mice were used in each group for the studies.