IMPACT OF EXTRACELLULAR VESICLES RELEASED BY HUMAN NEUTROPHILS TREATED WITH SHIGA TOXIN ON EPITHELIAL AND ENDOTHELIAL RENAL CELLS

MAIUMI SHIROMIZU C; KEITELMAN I; GOMEZ, F; VEREERTBRUGGHEN, A; VERA AGUILAR, DOUGLAS; PEREZ, P; ROSATO, M; RAMOS MV; JANCIC C; FEDERICO FUENTES; GALLETTI J; AMARAL, M; PALERMO, MARINA; SABBIONE F; TREVANI AS

Mar del Plata

Congreso; LXVII reunión anual de la sociedad argentina de investigación clínica (SAIC). LXX reunión anual de la sociedad argentina de inmunología (SAI) y 3er congreso franco-argentino de inmunología (FAIC). Reunión anual 2022 de la sociedad argentina de fisiología; 2022

Shiga toxin (Stx) producing Escherichia coli (STEC) is a non-invasivepathogen that colonises the intestine where it releases the Stxwhich can reach the blood stream and lead to the Haemolytic UremicSyndrome (HUS). In HUS neutrophilia is a typical sign and a poorprognosis factor. Previous studies suggest that Stx is transported totarget organs like kidneys, in extracellular vesicles (EV) generatedby blood cells. The aim of this study was to determine if neutrophils(N) produce EV in response to Stx (EV-Stx) and their impact on theviability and cytokine production by primary human glomerular endothelialcells (HGEC) and renal epithelial cells (HK-2 cells). HumanN (106) isolated from peripheral blood were treated with purifiedStx2 (100 ng/ml), heat-inactivated Stx2 (StxØ) or vehicle (C) for 4 hand EV released were isolated by differential centrifugation. By bothconfocal laser scanning microscopy (n=4) and detection of CD63expression by western blot (n=6), we determined that N release EVin all the conditions studied. By employing the VERO cell line susceptibleto Stx, we observed that EV-Stx but not EV-StxØ or EV-Csignificantly reduced cell viability (n=10; p