MURINE MACROPHAGES SURFACE N-GLYCANS PARTICIPATE IN YopP-MEDIATED IMMUNOPATHOGENESIS OF Yersinia enterocolitica INFECTIONS

GARAY, JUAN AGUSTÍN; SILVA JUAN EDUARDO; VARAS, JOSÉ; FUNES, SAMANTA CELESTE; JOFRE, BRENDA LUCILA; DI GENARO, MARÍA SILVIA; DAVICINO, ROBERTO CARLOS

Buenos Aires

Congreso; REUNIÓN CONJUNTA SAIC SAI&FAIC SAFIS 2022; 2022

Yersinia enterocolitica (Ye) outer membrane protein P (YopP) is ableto manipulate macrophages (MΦ) by inducing apoptosis and suppressing signaling molecules production. Although YopP has beenshown to interact in a carbohydrate-dependent manner with hostproteins, its interaction with glycans remains unclear. The aim ofour work is to explore the influence of MΦ surface N-glycans onthe immunomodulatory effects triggered by YopP. Murine peritonealMΦ were obtained by intraperitoneal lavage, plated at a 1x106 cells/mL and purified by incubation for 2 h at 37°C in DMEM medium inan atmosphere with 5% CO2. Then, MΦ were incubated with 5 IU/mL of PNGase F in DMEM medium without Fetal Bovine Serum for4 h (MΦd) and cultured overnight in supplemented DMEM medium. Subsequently, MΦ were infected with Ye serotype 0:8 (pYV+,WA-314) (Ye wt) or with Ye WA-314 deficient in YopP (pYV+, WA-CpYVNalrKanr) (Ye ∆yopP) at MOI 10:1. Then, MTT, lactate dehydrogenase (LDH), Griess and Urease assays were performed. Apoptosis was evaluated by flow cytometry using Annexin V-propidiumiodide, and TNF-α was measured by ELISA. Our results shows thatN-deglycosylation does not affect viability. Besides, we observed nosignificant apoptosis in Ye wt-infected MΦd compared to uninfectedcounterpart, suggesting a critical impact of N-glycans in the wellknown triggering role of Ye on apoptosis. On the other hand, weshowed that YopP has a dual effect: in MΦ inhibits NO production(p