Trophoblast cell invasion, glucose uptake and trophoblast-neutrophil interaction is impaired by LPS from Porphyromonas gingivalis

CALO, GUILLERMINA; PAPARINI, DANIEL; HAUK, VANESA C.; VOTA, DAIANA; RAMHORST, ROSANNA; LARA, BRENDA; MERECH, FÁTIMA; PÉREZ LEIRÓS, CLAUDIA

Virtual

Congreso; ASRI annual meeting; 2021

Problem Porphyromonas gingivalis (Pg) is 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 TLR4 and TLR2 mediated pathways. During placentation, trophoblast cells secrete cytokines and chemokines in order to interact with immune cells regulating and maintaining immune homeostasis. In fact, neutrophil activation is associated with poor placentation and severe pregnancy complications. Alterations in this interaction may lead to pregnancy complications. The aim of this study was to examine the effect of Pg-LPS on trophoblast cell function and to explore TLR4/TLR2 mediated mechanisms.Method of StudySwan-71 human trophoblastic cell line was treated with Pg-LPS (1-100ng/ml). Cytokine and chemokine expression was evaluated by RTqPCR or flow cytometry, glucose uptake by flow cytometry using the fluorescent analogue 2-NBDG, cell migration by wound-healing assay 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 treatment reduced cell migration and invasion (N=4 *P≤0.05). This effect was accompanied by a significant increase TIMP1, TIMP2 and TGF1 expression. We also found that Pg-LPS treatment reduced glucose uptake (P≤0.05) and decreased GLUT-1 expression. Pg-LPS treatment also impaired the balance of and decreased. Depending on the function studied TLR2 and/or TLR4 signalling pathways appeared differentially involved. Conditioned media from trophoblast cells with Pg-LPS increased neutrophil activation with higher release of ROS and lower apoptosis rate.ConclusionsIn summary, our results indicate that P. gingivalis lipopolysaccharide activation of TLR2 and TLR4 on trophoblast cells affects trophoblast cell invasion, cytokine expression and glucose uptake. leading to a deficient regulation of neutrophil proinflammatory profile This mechanism underlying Pg infection during placentation might contribute to the pathogenic effect of this bacteria on pregnancy outcome.