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

ROMINA S ALVAREZ; CAROLINA JANCIC; CRISTINA IBARRA; MARIA M AMARAL

Mar del Plata

Congreso; Reunión Conjunta SAIC SAI SAFIS 2018; 2018

SAIC-SAI-SAFIS

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 theHUS pathogenesis. To clarify the involvement of endothelial-epithelial crosstalk in the kidney toxins damages, previously, we developedan 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 (0.01ng/ml), SubAB(1ng/ml) or Stx2+SubAB, we had found a differential release of IL6, IL-8 and TNF-α between monocultures and cocultures, havingconsidered both compartments together. While IL-6, IL-8 and TNF-αrelease was increased on the coculture and HGEC monoculture byall treatments and Stx2, respectively, it was not modulated on HK-2monoculture by the toxins. In this work, we compared the releaseof these soluble mediators, measured in supernatants by ELISA,between HGEC and HK-2 monocultures respect to HGEC or HK-2coculture compartments. On the HGEC side, only Stx2 increasedIL-6 and IL-8 secretion. Surprisingly, on the HK-2 side, all treatmentscaused a significant increase in IL-6 and IL-8 release and statistically different from HK-2 monoculture. TNF-α was increased on bothHGEC and HK-2 side by all conditions (p