Crosstalk between human microvascular endothelial cells and tubular epithelial cells modulates pro-inflammatory response induced by Shiga type 2 and Subtilase toxins

JANCIC, CAROLINA C.; ALVAREZ, ROMINA S.; AMARAL, MARÍA MARTA; IBARRA, CRISTINA

Mar del Plata

Congreso; 63 Reunión Anual de la Sociedad Argentina de Investigación Clínica; 2018

SAI - SAIC

Shiga toxin (Stx) is considered the main virulence factor associated to the renal damage of the hemolytic uremic syndrome (HUS). However, Subtilase (SubAB) cytotoxin has also been related to the HUS pathogenesis. To clarify the involvement of endothelial-epithelial crosstalk in the kidney toxins damages, previously, we developed an in vitro model of renal proximal tubule by microvascular endothelial cells (HGEC) and tubular epithelial cells (HK-2) in coculture (HGEC/HK-2). After 24h of incubation with Stx2 (0.01 ng/ml), SubAB (1 ng/ml) or Stx2+SubAB, we had found a differential release of IL-6, IL-8 and TNF-α between monocultures and cocultures, having considered both compartments together. While IL-6, IL-8 and TNF-α release was increased on the coculture and HGEC monoculture by all treatments and Stx2, respectively, it was not modulated on HK-2 monoculture by the toxins. In this work, we compared the release of these soluble mediators, measured in supernatants by ELISA, between HGEC and HK-2 monocultures respect to HGEC or HK-2 coculture compartments. On the HGEC side, only Stx2 increased IL-6 and IL-8 secretion. Surprisingly, on the HK-2 side, all treatments caused a significant increase in IL-6 and IL-8 release and statistically different from HK-2 monoculture. TNF-α was increased on both HGEC and HK-2 side by all conditions (p