INVESTIGADORES
VOTA daiana Marina
congresos y reuniones científicas
Título:
LPS from Porphyromonas gingivalis impairs trophoblast function and trophoblast-neutrophil interaction
Autor/es:
LARA, BRENDA; CALO, GUILLERMINA; VOTA, DAIANA; MERECH, FÁTIMA; PAPARINI, DANIEL ; RAMHORST, ROSANNA; PÉREZ LEIRÓS, CLAUDIA; HAUK, VANESA
Reunión:
Simposio; International Symposium of Reproductive Health; 2021
Resumen:
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) is the main virulence factor of Pg. 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. Objectives The aim of this study was to examine the effect of Pg-LPS on trophoblast cell function and to explore TLR4/TLR2 mediated mechanisms. Methods Swan-71 human trophoblastic cell line was treated with Pg-LPS. Cytokine and chemokine expression were evaluated by RTqPCR or flow cytometry, 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. Results Pg-LPS treatment reduced cell migration and invasion (N=4 *P≤0.05). 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 cytokines and chemokines. 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. Conclusions In 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.