Murine peritoneal macrophages surface n-glycans regulate respiratory burst through ERK 1/2 and p38 MAPKS pathway

JUAN AGUSTÍN GARAY; JUAN EDUARDO SILVA; BRENDA JOFRÉ; MARÍA SILVIA DI GENARO, ; ROBERTO CARLOS DAVICINO

Mar del Plata

Congreso; Reunión Conjunta SAIC SAI&FAIC SAFIS 2022; 2022

Sociedades científicas SAIC, SAI&FAIC, SAFIS

Recent studies have explored the role of surface N-glycans in neutrophil effector functions, however there is little information regarding macrophages (MΦ). Thus, in the present work, murine peritoneal MΦ treated with PNGase F were used in order to study N-glycans role on respiratory burst. MΦ were obtained through peritoneal lavage with saline solution and adjusted at 1x106 cells/ml, then purified by incubation for 2 h at 37°C in DMEM medium supplemented with 10% fetal bovine serum (FBS) in an atmosphere with 5% CO2. Then, MΦ were incubated with 5 IU of PNGase F in DMEM medium without FBS for 4 h, then, washed and cultured overnight in supplemented DMEM medium. After N-deglycosylation, NBT assay was performed and Cd2+ (NADPH oxidase inhibitor), malonate, and barbital (mitochondrial complex II and I inhibitors, respectively) were used. Flow cytometry analysis was performed with the DHR-PMA assay in presence and absence of ERK1/2, p38, and JNK inhibitors. The expression of CD115, CD11b, F4/80 and Ly6C was also examined. Finally, we evaluated phagocytosis using Yersinia enterocolitica (Ye)-GFP assay. We have found that N-deglycosylation increases the respiratory burst according to NBT test (p