Sexual dimorphism in the immunometabolic profile of Hofbauer cells

PAPARINI D, GRASSO E, AGUILERA F, LARA B, ; ARSLANIAN MA, LELLA V,; HAUK V , MERECH M, SCHUSTER C, GORI S, ; MELLER C; MARTÍ M, RAMHORST R , VOTA D, ; PEREZ LEIROS C

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

Aim: Metabolic reprogramming of macrophages is associated to functionalpolarization upon environmental stimuli. In normal pregnancy, monocytesrecruited to the pregnant uterus differentiate into M2-like decidual macrophagesunder the control of extravillous trophoblast cells. However, the immunometabolicprofiling of human maternal circulating monocytes at early pregnancy andwhether the trophoblast has a conditioning role have not been reported so far.We aim to characterize the metabolic pathways active in circulating monocytesfrom pregnant women at 16-20 weeks in basal and ex vivo LPS-stimulatedconditions. Methods and Results: Peripheral blood mononuclear cells (PBMC)were isolated from fertile non-pregnant and 16-20 weeks pregnant women byFicoll-Paque and stimulated or not with 100 ng/ml LPS from E. coli. CD14+ cellmetabolism 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 CD14antibody staining was followed by intracellular anti-puromycin staining. Glucoseand long chain polyunsaturated fatty acids (LCPUFA) uptake was assessed byflow cytometry with D-glucose fluorescent analog (2-NBDG) or Bodipy FL-C12,respectively. IL-1β and IL-10 secretion was measured by ELISA and efferocytosisof apoptotic autologous neutrophils stained with CFSE was quantified by flowcytometry. Results: Monocytes from pregnant vs. non-pregnant women showedhigher glucose dependency (p