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

MERECH F, CALO G, RIOS D, DOGA L, DERAMO L, LARA B, PAPARINI D, SQUASSI A, HAUK V, RAMHORST R, ; ARGÜELLO RJ; PEREZ LEIROS C; VOTA D

Congreso; Reunion Anual de la Sociedad Argentina de Inmunologia; 2023

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