Prereceptor amplification of corticosterone by splenic DC subsets. (A) Estimated 11βHSD1 reductase activity of CD11c⁺ DCs and CD4 cells (n = 4 per group; 2 independent experiments). (B) Corticosterone generation after 18 hours of pDC or CD11c⁺ DC culture in the presence or absence of A at 10⁻⁷ M (n = 10 to 11 per group). (C) Corticosterone in pDCs (n = 4 to 10 per group) or pooled CD11c⁺ DCs (n = 3 to 11 per group) from spleens of H6pdh^−/− mice or H6pdh^+/+ littermate controls. Data in Figure 2B-C are derived from 3 or more independent experiments. (D) Wild-type B6 mice received phosphate-buffered saline (PBS) or the indicated TLR agonists by intraperitoneal injections (CpG, 150 µg; poly I:C, 100 µg; LPS, 10 µg; Flagellin, 5 µg) 18 hours prior to pDC, CD8α⁺ DC, and CD8α^– DC isolation from the spleen (n = 3 per group per condition; data from 2 independent experiments). Graph shows mean ± standard deviation (SD) corticosterone generation in each experimental group after exposure to A at 10⁻⁶ M (rather than 10⁻⁷ M as in Figure 2B-C). Higher concentrations of input A were used to permit detection of B produced by CD8α^– DCs. (E) Wild-type B6 mice received PBS or 50 µg anti-CD40 agonist antibody 18 hours prior to isolation of DC subsets from the spleen (n = 3 per group per condition; data from 2 independent experiments). Graph shows mean ± SD corticosterone generation after exposure to A at 10⁻⁶ M in each experimental group.