Cytotoxic effect of Shiga toxin-1 on human proximal tubule cells.
BACKGROUND
Cytolytic Shiga toxins (Stx) are believed to be largely responsible for renal damage in post-diarrheal hemolytic-uremic syndrome (D + HUS). Despite the general belief that endothelial cells are the primary target of Stx, there is evidence that proximal tubules may be a site of toxin action. We hypothesized that cultured proximal tubular cells are sensitive to the cytotoxic effects of Stx.


METHODS
Cultured human proximal tubular cells were exposed to Stx-1 in the presence and absence of a variety of inflammatory factors likely to be elevated in the kidney or serum of patients with D + HUS. Cell survival, protein synthesis, total cell levels and synthesis of Stx receptors (GB3), and Stx binding were measured.


RESULTS
Proximal tubules were extremely sensitive to the cytotoxic effect of Stx-1 with an LD50 at least equal to, if not less than, that seen with Vero cells. Interleukin-1 (IL-1), lipopolysaccharide (LPS), and butyrate (but not tumor necrosis factor or interleukin-6) up-regulated proximal tubule sensitivity to Stx-1. IL-1 increased Stx-1 binding, but did not alter total cell levels or synthesis of GB3, the glycosphingolipid receptor for Stx-1. In contrast, LPS and butyrate, despite increasing Stx-1 sensitivity, had no effect on Stx-1 binding.


CONCLUSIONS
These studies indicate that proximal tubules are exquisitely sensitive to Stx-1 cytotoxicity and that inflammatory factors can increase toxin responsiveness through a variety of mechanisms. It is suggested that proximal tubules may be an important early target of Stx-1 action in D + HUS.
Hughes
A K
AK
Veterans Affairs Medical Center, Salt Lake City, Utah, USA.
Stricklett
P K
PK
Kohan
D E
DE
D001427:Bacterial Toxins / Q000633:toxicity*; D002478:Cells, Cultured; D003513:Cycloheximide / Q000494:pharmacology; D016207:Cytokines / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007687:Kidney Tubules, Proximal / Q000187:drug effects* / Q000473:pathology; D008070:Lipopolysaccharides / Q000494:pharmacology; D014176:Protein Biosynthesis; D022603:Shiga Toxins; D014281:Trihexosylceramides / Q000032:analysis
D001427:Bacterial Toxins; D016207:Cytokines; D008070:Lipopolysaccharides; D022603:Shiga Toxins; D014281:Trihexosylceramides; C018549:globotriaosylceramide; D003513:Cycloheximide