E2 treatment enhances IFN-α production by TLR-9 ligand-stimulated pDCs in postmenopausal women. (A) PBMCs purified from whole blood were cultured at 5 × 105 cells/well in the presence of indicated concentrations of CpG-2216. IFN-α concentration was measured by ELISA in 24-hour culture supernatants. The IFN-α range measured for 1 μg/mL of CpG was from 21 pg/mL to 2914 pg/mL. Data were normalized to the number of pDCs present in PBMCs, as determined by flow cytometry. Data from 8 representative subjects are shown, before (dashed lines) or after E2 treatment (solid line). Numbers in parentheses show the fold increase of IFN-α production as calculated in panel D. (B-C) Normalized IFN-α production before (B) and after (C) E2 treatment was calculated by summing the concentrations of IFN-α measured in response to the different concentrations of CpG as shown in panel A (n = 23). Dot plot panels show the correspondence between the values for each patient in samples S1 versus S2 (B) and S2 versus S3 (C). (D) Data from samples S2 and S3 were expressed as fold increase of normalized IFN-α production relative to the S1 and mean S1-S2 values, respectively. Individual data are shown, and horizontal bars indicate mean values. P values were determined using the Wilcoxon signed rank test.
