LPS from porphyromonas gingivalis impairs trophoblast cell function and immune-trophoblast interaction.

V. HAUK , G. CALO, M. ROMÁN MUÑOZ, D. VOTA, F.I. MERECH, D. PAPARINI, R. RAMHORST, C. PEREZ LEIROS

Buenos Aires

Congreso; International Federation of Placenta Association meeting 2019; 2019

ObjectivesPorphyromonas gingivalis (Pg), an important pathogen of periodontal disease, has been implicated in adverse pregnancy outcome although the mechanisms involved are still unclear. Lipopolysaccharide from Porphyromonas gingivalis (Pg-LPS), the main virulence factor of Pg, differs from other bacterial LPS in its structure and function: due to the presence of lipoproteins, Pg-LPS activates not only TLR4 but also TLR2 mediated pathways. The aim of this study was to examine the effect of Pg-LPS on trophoblast cell function and trophoblast-neutrophil interaction and to explore TLR4/TLR2 mediated mechanisms.MethodsSwan-71 human trophoblastic cell line was treated with Pg-LPS (10ng/ml). Cytokine and chemokine expression was evaluated by RTqPCR, glucose uptake by flow cytometry using the fluorescent analogue 2-NBDG and cell invasion assessed in Matrigel-covered transwells Peripheral blood neutrophils were purified from healthy donors and cultured with conditioned media of trophoblast cells (TbCM) treated or not with LPS (PgLPS-CM); apoptosis was determined by fluorescence microscopy and CD11b and reactive oxygen species (ROS) were evaluated by flow cytometry. ResultsPg-LPS altered trophoblast function: It reduced cell invasion, impaired pro-inflammatory/ antiinflamatory cytokine expression and decreased glucose uptake (P≤0.05). Depending on the function studied TLR2 and/or TLR4 signalling pathways appeared differentially involved. PgLPS-CM increased neutrophil activation with higher CD11b expression and higher release of ROS (X±SE: basal Neutrophil: 13930±2830; Neutrophil+TbCM: 10721±2298 Neutrophil+PgLPS-CM: 19816±3543 P