PGE2 promotes BM-DC chemotactic migration through Matrigel in an MMP-9–dependent manner. (A) CD11c+ DCs were treated with IFN-α + TNF-α with or without PGE2 (10−6 M), in the presence or absence of MMP-9 inhibitor I for 48 hours. DCs (105 cells in 0.1 mL) were placed in the upper Transwell chambers precoated with Matrigel (70 μg). The bottom chambers were filled with serum-free medium with or without CCL19 (100 ng/mL). Following incubation at 37°C for 4 hours, the migrating cells were collected from the lower chambers and counted by FACS (60-second counts). (B,C) DCs generated from wild-type (WT) FBV/NJ or MMP-9–deficient mice were treated with IFN-α + TNF-α with or without PGE2 (10−6 M) for 48 hours. The amounts of pro–MMP-9 (B) and total MMP-2 (C) were quantified by ELISA. (D) DCs generated from wild-type (WT) FBV/NJ or MMP-9–deficient mice were treated with IFN-a + TNF-α with or without PGE2 (10−6 M), in the presence or absence of the MMP-9 inhibitor I (10−6 M) or MMP-2 inhibitor I (10−5 M) for 48 hours. The cells were placed in the Transwell upper chambers precoated with Matrigel. The bottom chambers were filled with serum-free medium with or without CCL19 (100 ng/mL). After 5 hours of incubation at 37°C, the migrating cells were collected from the lower chambers and counted by FACS (60-second counts). Data are representative of 3 independent experiments.
