Figure 5. Trophoblast cells selectively reduce chemokine production after the interaction with maternal monocytes in the presence of PAMPs stimuli.

Swan-71 cell line was cultured with LPS (10 µg/ml), PGN (10 µg/ml), or poly [I:C] (10 µg/ml) in a polystyrene plate in the absence or presence of 0.4 um pore-insert with CD14+ cells. After 24 hours, Swan-71 cells were recovered from the lower compartments and the expression of CCL5, CXCL8 and CCL2 (A) was evaluated by RT-PCR and normalized to GAPDH expression. Results are expressed as arbitrary units (a.u) (mean ± SEM) from 3 independent experiments using different fertile women (*p<0.05, Friedman test followed by Dunn’s post-test). (B) Representative amplification bands from 3 independent experiments. (C) Migration assays were performed in transwell system. Condition media from Swan-71 cell cultured in the absence or presence of poly [I:C] (10 µg/ml) was used as attractant in the lower compartment pre-treated or not with anti-CCL5 neutralizing Ab and 8 µm-pore inserts containing the isolated monocytes were added (upper compartment). After 24 hours, cells were recovered from the lower and upper compartments and the frequency of CD14+ cells were quantified by FACS analysis. Results are expressed as fold increase of CD14+ migration relative to basal condition media migration (mean ± SEM) from 3 independent migration assays (*p<0.05, Friedman test followed by Dunn’s post-test).