Early pregnancy reprograms maternal circulating monocyte metabolismo to higher glucose dependency associated to increased efferocytosis

MERECH, FÁTIMA; CALO, GUILLERMINA; RIOS, DAIANA; DOGA, LUCIANA; DERAMO, LUCIANA; PAPARINI, DANIEL E.; LARA, BRENDA; SQUASSI, ALDO; HAUK, VANESA; RAMHORST, ROSANNA; ARGÜELLO, RAFAEL J.; PÉREZ LEIRÓS, CLAUDIA; VOTA, DAIANA M.

San Luis

Congreso; LXXI REUNION ANUAL DE LA SOCIEDAD ARGENTINA DE INMUNOLOGIA (SAI); 2023

SAI

Aim: Metabolic reprogramming of macrophages is associated to functional polarization upon environmental stimuli. In normal pregnancy, monocytes recruited to the pregnant uterus differentiate into M2-like decidual macrophages under the control of extravillous trophoblast cells. However, the immunometabolic profiling of human maternal circulating monocytes at early pregnancy and whether the trophoblast has a conditioning role have not been reported so far. We aim to characterize the metabolic pathways active in circulating monocytes from pregnant women at 16-20 weeks in basal and ex vivo LPS-stimulated conditions. Methods and Results: Peripheral blood mononuclear cells (PBMC) were isolated from fertile non-pregnant and 16-20 week pregnant women by Ficoll-Paque and stimulated or not with 100 ng/ml LPS from E. coli. CD14+ cell metabolism was immediately analyzed using SCENITHTM by flow cytometry. Briefly, metabolic inhibitors (2-Deoxy-D-glucose, Oligomycin or their combination) were added to PBMC for 15 min. Then puromicyn was added 30 min and CD14 antibody staining was followed by intracellular anti-puromycin staining. Glucose and long chain polyunsaturated fatty acids (LCPUFA) uptake was assessed by flow cytometry with D-glucose fluorescent analog (2-NBDG) or Bodipy FL-C12, respectively. IL-1β and IL-10 secretion was measured by ELISA and efferocytosis of apoptotic autologous neutrophils stained with CFSE was quantified by flow cytometry. Results: Monocytes from pregnant vs. non-pregnant women showed higher glucose dependency (p