CONICET | Buscador de Institutos y Recursos Humanos

During placentation, trophoblast cells interact and secrete cytokine in order to regulate and maintain immune homeostasis. Changes or defects in this interaction may lead to pregnancy complications. In fact neutrophil activation is associated with poor placentation and severe pregnancy complications. Porphyromonas gingivalis (Pg) is an important pathogen of periodontal disease that has been implicated in adverse pregnancy outcome although the mechanisms involved are still unclear. Pg-LPS is the most important virulence factor of Pg and activates both TLR4 and TLR2.The aim of this work was to evaluate the effect of conditioned media of trophoblast cells primed with Pg-LPS on neutrophil function. Human trophoblastic cell line Swan-71 was treated with Pg-LPS(10ng/ml) or Pg-LPS ultrapure, variant that only activates TLR4. Cytokine expression was evaluated by RTqPCR, flow cytomery and ELISA. Peripheral blood neutrophils (Neu) and mononuclear cells(PBMCs) were purified from healthy donors and cultured with conditioned media from trophoblast cells (TbCM) treated or not with LPS (PgLPS-CM) or LPS ultrapure (PgLPSultra-CM). Apoptosis andreactive oxygen species (ROS) were evaluated by flow cytometry Regulatory T cell induction was evaluated by flow cytometry. Pg-LPS ultrapure increasde the expression of TNFa, IL6 and IL1brespect to basal while Pg-LPS had no effect. In line with this result, Pg-LPS-CM had no effect on neutrophil activation whereas PgLPSultra-CM increased neutrophil activation with higher release ofROS and decreased apoptosis rate (p≤0.05). Neutrophils exposed to TbCM induced FOXP3 expression on CD4+ T cell after 48h of coculture with PBMCs. This induction was diminished when neutrophil were preconditioned with PgLPS-CM or PgLPSultra-CM (P