Intracellular edema produced by Shiga toxin type 2 in human glomerular endothelial cells: Role of aquaporins and Gb3 receptor

GOMEZ, FERNANDO; REPETTI, JULIETA; GIRON REYES, CLAUDIO DANIEL; SACERDOTI, FLAVIA; PRESTA, AGOSTINA; IBARRA, CRISTINA; MARTINEZ, NORA; DAMIANO, ALICIA E.; DI GIUSTO, GISELA; AMARAL M MARTA

Banff

Congreso; 11th INTERNATIONAL SYMPOSIUM on Shiga Toxin (Verocyotoxin) Producing Escherichia coli Infections; 2023

VTEC

Hemolytic Uremic Syndrome (HUS) related to Shiga toxin – producing Escherichia coli (STEC) is the major cause of acute renal injury in the pediatric age group in Argentina. Previously, we have demonstrated that Shiga toxin type 2 (Stx2) inhibits the net water transport (Jw) across monolayers of human glomerular endothelial cells (HGEC), possibly as a consequence of direct alterations in the Jw mechanisms. In addition, we figured out that Stx2 damages HGEC by inducing swelling and detachment. In this work, we analyzed changes in the cell area  and in the cell volume of HGEC      monolayers exposed to Stx2 in the presence or not of aquaporins (AQPs) inhibitors as mercury chloride (HgCl2) and tetraethylammonium (TEA) or an inhibitor of Stx2 receptor (Gb3) synthesis, Eliglustat (EG). In addition, we assayed the expression of both AQP1 and AQP4 in the presence or not of Stx2. For cell area studies, HGEC were grown on 12 well plates and pretreated with HgCl2 (10 μM) or TEA (100 μM) for 30 minutes, or with EG (10 μM) for 24 hours and under isotonic conditions. Then, HGEC were incubated with Stx2 (5 ng/ml) for an additional 40 minutes and then analyzed by light microscopy. Our results point out that, after Stx2 incubation, a significant increase (42%) in the cell area of HGEC (p