INTRODUCTION: Bacterial vaginosis (BV), characterised by an imbalance of the vaginal microbiome and genital inflammation, is a risk factor for transmitting and contracting human immunodeficiency virus (HIV). Lactic acid (LA), a vaginal microbiota metabolite (VMB) produced by lactobacilli, is elevated during vaginal eubiosis and elicits an anti-inflammatory effect on cervicovaginal epithelial cells that could protect against HIV. In contrast, BV is characterised by a dramatic depletion of LA and an increase in VMB's comprising short-chain fatty acids (SCFAs) and succinic acid (SA). SCFAs elicit a proinflammatory response from PBMCs; however the impact of SCFAs on cervicovaginal epithelial cells is unknown.
METHODS: Monolayers of ectocervical cells (Ect1/E6E7) were treated with combinations of VMB to mimic eubiotic and BV conditions in the absence or presence of TLR agonists (LPS, Poly I:C and Pam3CSK4) to simulate pathogen challenge. Cytokines and chemokines in the supernatant were quantified using a Luminex multiplex assay.
RESULTS: VMB representing BV conditions had no effect on the pro- and anti-inflammatory response of Ect1 cells. In the presence of TLR agonist stimulation, BV VMB had little effect on dampening cytokine/chemokine responses compared to eubiotic VMB. Increased production of the anti-inflammatory cytokine IL-1RA was observed after eubiotic VMB treatment both with and without TLR agonist stimulation, but not with VMB representing BV conditions. Increased secretion of pro-inflammatory cytokines IL-6 and IL-8 by TLR agonists was decreased under only eubiotic VMB conditions. Similarly, elevated IP-10, TNF, RANTES and MIP3α levels elicited by poly I:C treatment was only dampened by eubiotic VMB
CONCLUSION: In contrast to the anti-inflammatory effect of eubiotic VMB due to LA, VMB representing BV conditions failed to elicit immune modulatory effects on Ect1 cells and had little impact on dampening pro-inflammatory immune mediators elicited by TLR agonists consistent with the pro-inflammatory milieu associated with BV and increased HIV risk.