Pregnancy shapes immunometabolic reprogramming of maternal circulating monocytes.

. F MERECH, G REYSCHER, V HAUK, G CALO, S NOVOA, L DOGA, L D´ERAMO, P FABBIANO, A SQUASSI, R RAMHORST, R ARGÜELLO, C PÉREZ LEIROS, D VOTA

Congreso; Reunión Anual Conjunta SAIC-SAI-SAFIS; 2022

Metabolic reprogramming of macrophages is associated to functional polarization upon environmental stimuli. Decidual macrophagesdisplay high plasticity to face opposing demands such as toleranceto embryonic antigens and efficient control of pathogens. In normalpregnancy, trophoblast cells drive macrophages to alternative M2phenotypes. So far, the immunometabolic profile of maternal monocytes during pregnancy and the effect of infectious stimuli werenot characterized. Objective: To investigate the metabolic profile ofmonocytes at early pregnancy and the effect of inflammatory bacterial stimuli. Methods and Results: Peripheral blood mononuclearcells (PBMC) were isolated from fertile non-pregnant and 16-20week pregnant women by Ficoll-Paque and stimulated with 100ng/ml LPS from E. coli or periodontal pathogens. CD14+ cell metabolism was analyzed by flow cytometry after SCENITH protocol.Briefly, metabolic inhibitors (2-Deoxy-D-glucose, Oligomycin or theircombination) were added to PBMC for 15 min. Then puromicyn wasadded 30 min and CD14 antibody staining was followed by intracellular antipuromycin staining. Glucose and long chain polyunsaturated fatty acids (LCPUFA) uptake was assessed by flow cytometrywith D-glucose fluorescent analog (2-NBDG) or Bodipy FL-C12,respectively. CD14+cells from pregnant vs. non-pregnant womenshowed higher glucose dependency and glycolytic capacity alongwith decreased mitochondrial dependency and fatty acid & aminoacid oxidation capacity (p