Shiga toxin 1-induced inflammatory response in lipopolysaccharide-sensitized astrocytes is mediated by endogenous tumor necrosis factor alpha.
Hemolytic-uremic syndrome (HUS) is generally caused by Shiga toxin (Stx)-producing Escherichia coli. Endothelial dysfunction mediated by Stx is a central aspect in HUS development. However, inflammatory mediators such as bacterial lipopolysaccharide (LPS) and polymorphonuclear neutrophils (PMN) contribute to HUS pathophysiology by potentiating Stx effects. Acute renal failure is the main feature of HUS, but in severe cases, patients can develop neurological complications, which are usually associated with death. Although the mechanisms of neurological damage remain uncertain, alterations of the blood-brain barrier associated with brain endothelial injury is clear. Astrocytes (ASTs) are the most abundant inflammatory cells of the brain that modulate the normal function of brain endothelium and neurons. The aim of this study was to evaluate the effects of Stx type 1 (Stx1) alone or in combination with LPS in ASTs. Although Stx1 induced a weak inflammatory response, pretreatment with LPS sensitized ASTs to Stx1-mediated effects. Moreover, LPS increased the level of expression of the Stx receptor and its internalization. An early inflammatory response, characterized by the release of tumor necrosis factor alpha (TNF-alpha) and nitric oxide and PMN-chemoattractant activity, was induced by Stx1 in LPS-sensitized ASTs, whereas activation, evidenced by higher levels of glial fibrillary acid protein and cell death, was induced later. Furthermore, increased adhesion and PMN-mediated cytotoxicity were observed after Stx1 treatment in LPS-sensitized ASTs. These effects were dependent on NF-kappaB activation or AST-derived TNF-alpha. Our results suggest that TNF-alpha is a pivotal effector molecule that amplifies Stx1 effects on LPS-sensitized ASTs, contributing to brain inflammation and leading to endothelial and neuronal injury.
Landoni
Verónica I
VI
Departamento de Immunología, Academia Nacional de Medicina, Pacheco de Melo 3081, 1425 Buenos Aires, Argentina. vilandoni@yahoo.com.ar
de Campos-Nebel
Marcelo
M
Schierloh
Pablo
P
Calatayud
Cecilia
C
Fernandez
Gabriela C
GC
Ramos
M Victoria
MV
Rearte
Bárbara
B
Palermo
Marina S
MS
Isturiz
Martín A
MA
D000818:Animals; D001253:Astrocytes / Q000187:drug effects* / Q000276:immunology*; D002478:Cells, Cultured; D004926:Escherichia coli / Q000472:pathogenicity; D018836:Inflammation Mediators / Q000378:metabolism; D008070:Lipopolysaccharides / Q000276:immunology*; D009569:Nitric Oxide / Q000378:metabolism; D051381:Rats; D022622:Shiga Toxin 1 / Q000276:immunology* / Q000633:toxicity*; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*
D018836:Inflammation Mediators; D008070:Lipopolysaccharides; D022622:Shiga Toxin 1; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide