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Synergic Activation of Toll-Like Receptor (TLR) 2/6 and 9 in Response to Ureaplasma parvum & urealyticum in Human Amniotic Epithelial Cells

Figure 2

TLR2/6 and TLR9-dependent cytokine secretion in response to Ureaplasma serovars.

Human amniotic epithelial cells (A, B) were either not stimulated (white bar charts) or stimulated with Ureaplasma serovars for 2 hours. The cells were fixed and permeabilised, followed by antibody staining against the particular TLR molecule, and incubation with the appropriate secondary antibody conjugated to FITC. Fluorescence was detected using a FACSCalibur (BectonDickinson The data presented is the mean of three independent experiments. HEK-293 cells (B) transfected with TLR1, TLR2, TLR2/6, TLR4, TLR7 and TLR9 were either not incubated (white bar charts) or incubated with with Ureaplasma (1×108 bacteria/ml to 1×107 cells/ml) parvum SV3 (black barcharts), U. parvum SV14 (grey barcharts), U. urealyticum SV2 (stripped bar charts), or MBA (1 µg/ml) for 2 h. Control cultures were stimulated with known TLR2, TLR4, TLR7 or TLR9 ligands. The supernatants were harvested and assayed for IL-6 content using the Cytometric Bead Array (CBA) system (Becton Dickinson). Fluorescence was detected using a FACSCalibur (BectonDickinson). The data represents the mean ± SD of three independent experiments. Asterisks indicate statistically significant (p<0.05) increase in expression (A,B) or IL-6 secretion (C) compared to corresponding unstimulated controls.

Figure 2

doi: https://doi.org/10.1371/journal.pone.0061199.g002