Influence of communication between human renal endothelial and epithelial cells on pro-inflammatory mediators induced by Escherichia coli Shiga type 2 and Subtilase toxins

CRISTINA IBARRA; JAMES C PATON; HORACIO A REPETTO; CAROLINA JANCIC; ROMINA S ALVAREZ; MARÍA M AMARAL; ADRIENNE W PATON; CAROLINA M SHIROMIZU

Florencia

Simposio; 10th VTEC International Symposium on Shiga Toxin (Verocytotoxin) Producing Escherichia coli Infections; 2018

IntroductionHemolytic Uremic Syndrome associated to Shiga toxin (Stx)-producing E. coli infection is the most common cause of acute renal failure (ARF) in children in Argentina. Stx type 2 (Stx2) damages human renal microvascular endothelial cells (HGEC) and tubular epithelial cells (HK-2) and also induces a broad inflammatory response. Subtilase cytotoxin (SubAB) may also contribute to this pathology. We developed a HGEC/HK-2 coculture as a model of renal proximal tubule to clarify the involvement of the endothelial-epithelial cross talk in the kidney damage caused by toxins. In this work, we evaluated the response of selected pro-inflammatory mediators released by HGEC, HK-2 and HGEC/HK-2 exposed to Stx2, SubAB or Stx2+SubAB.MethodsHGEC, HK-2 and HGEC/HK-2 were incubated with Stx2 (0.01ng/ml), SubAB (1ng/ml) or Stx2+SubAB [(0.01+1) ng/ml] for 24 h. Culture supernatants were collected and IL-6, IL-8 and TNF-α were quantified by ELISA. Stx2, SubAB, Stx2+SubAB vs. Ctrl, p