Figure 3. Monocytes participate in the inhibition of IFN-α production in PBMCs by HBsAg.

(A) Magnetic bead-isolated monocytes and pDCs were co-cultured at varying ratios in RPMI 1640 medium supplemented with IL-3 (10 ng/ml) in 96-well plates. The pDCs (1×10⁴ per well) number was constant, and of the ratio of monocytes varied (0, 2, 5, 10, 20 fold). The cells were co-cultured and treated in the same manner as in 2B. The IFN-α production was determined by ELISA. The data are representative of at least three independent experiments. (B) The efficiency of CD14+ monocytes depletion. Monocytes were depleted using a CD14+ monocytes positive selection kit. The positive fraction was removed, and the negative fraction was collected. The purity of the depletion was determined by FACS analysis. (C) Total PBMCs and CD14+ monocyte-depleted PBMCs were treated in the same fashion as in 2B. IFN-α production was determined by ELISA. The data are representative of at least three independent experiments.