MURINE PERITONEAL MACROPHAGES SURFACE N-GLYCANS REGULATE RESPIRATORY BURST THROUGH ERK 1/2 AND P38 MAPKs PATHWAY

GARAY, JUAN AGUSTIN; JUAN SILVA; JOFRÉ B; MARÍA S. DI GENARO; DAVICINO RC

Mar del Plata

Congreso; Sociedad Argentina de Inmunología; 2022

SAI

Recent studies have explored the role of surface N-glycans in neutrophileffector functions, however there is little information regardingmacrophages (MΦ). Thus, in the present work, murine peritonealMΦ treated with PNGase F were used in order to study N-glycansrole on respiratory burst. MΦ were obtained through peritoneal lavagewith saline solution and adjusted at 1x106 cells/ml, then purifiedby incubation for 2 h at 37°C in DMEM medium supplementedwith 10% fetal bovine serum (FBS) in an atmosphere with 5% CO2.Then, MΦ were incubated with 5 IU of PNGase F in DMEM mediumwithout FBS for 4 h, then, washed and cultured overnight in supplementedDMEM medium. After deglycosylation, NBT assay was performedand Cd2+ (NADPH oxidase inhibitor), malonate, or barbital(mitochondrial complex II and I inhibitors, respectively) were used.Flow cytometry analysis was performed with the DHR-PMA assayin presence and absence of ERK1/2, p38, or JNK inhibitors. Theexpression 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 increasesthe respiratory burst according to NBT test (p