Figure 2.

A BV vaginal microbiota metabolite combination does not alter the production of immune mediators from Ect cells treated in a transwell system. Ect cells seeded in transwells were apically treated with a mixture of vaginal microbiota metabolites at pH 7 recapitulating bacterial vaginosis (BV; 6 mM L-LA, 100 mM acetic acid, 20 mM succinic acid, 4 mM propionic acid, and 4 mM butyric acid) in the absence or presence of TLR agonists PIC (TLR3) and Pam (TLR1/2) as indicated. Cells were stimulated for 1 h and supernatant collected from the apical compartment after an additional 18 h of culture. Production of IL-1RA (A), IL-6 (B), IL-8 (C), IP-10 (D), RANTES (E), and MIP-3α (F) was quantified using a luminex multiplex assay. Graphs show mean and standard error of the mean from n = 4 independent assays. Mann-Whitney tests were used to estimate statistical significance between treatment groups. + denotes a significant difference (p < 0.05) as compared to untreated (UT) cells. TLR, toll-like receptor; PIC, polyinosinic:polycytidylic acid; Pam, Pam3CSK4.