title: Shiga toxin 2 reduces complement inhibitor CD59 expression on human renal tubular epithelial and glomerular endothelial cells.
abstract: Infections with enterohemorrhagic Escherichia coli (EHEC) are a primary cause of hemolytic-uremic syndrome (HUS). Recently, Shiga toxin 2 (Stx2), the major virulence factor of EHEC, was reported to interact with complement, implying that the latter is involved in the pathogenesis of EHEC-induced HUS. The aim of the present study was to investigate the effect of Stx2 on the expression of membrane-bound complement regulators CD46, CD55, and CD59 on proximal tubular epithelial (HK-2) and glomerular endothelial (GEnC) cells derived from human kidney cells that are involved in HUS. Incubation with Stx2 did not influence the amount of CD46 or CD55 on the surface of HK-2 and GEnC cells, as determined by fluorescence-activated cell sorter analysis. In contrast, CD59 was significantly reduced by half on GEnC cells, but the reduction on HK-2 cells was less pronounced. With increasing amounts of Stx2, reduction of CD59 also reached significance in HK-2 cells. Enzyme-linked immunosorbent assay analyses showed that CD59 was not present in the supernatant of Stx2-treated cells, implying that CD59 reduction was not caused by cleavage from the cell surface. In fact, reverse transcription-quantitative PCR analyses showed downregulation of CD59 mRNA as the likely reason for CD59 cell surface reduction. In addition, a significant increase in terminal complement complex deposition on HK-2 cells was observed after treatment with Stx2, as a possible consequence of CD59 downregulation. In summary, Stx2 downregulates CD59 mRNA and protein levels on tubular epithelial and glomerular endothelial cells, and this downregulation likely contributes to complement activation and kidney destruction in EHEC-associated HUS.
authors.affiliation: Division of Hygiene and Medical Microbiology, Innsbruck Medical University, Innsbruck, Austria.
authors.forename: Silvia
authors.initials: S
authors.lastname: Ehrlenbach
authors.forename: Alejandra
authors.initials: A
authors.lastname: Rosales
authors.forename: Wilfried
authors.initials: W
authors.lastname: Posch
authors.forename: Doris
authors.initials: D
authors.lastname: Wilflingseder
authors.forename: Martin
authors.initials: M
authors.lastname: Hermann
authors.forename: Jens
authors.initials: J
authors.lastname: Brockmeyer
authors.forename: Helge
authors.initials: H
authors.lastname: Karch
authors.forename: Simon C
authors.initials: SC
authors.lastname: Satchell
authors.forename: Reinhard
authors.initials: R
authors.lastname: Würzner
authors.forename: Dorothea
authors.initials: D
authors.lastname: Orth-Höller
mesh_terms: D018957:CD59 Antigens / Q000378:metabolism*; D002460:Cell Line; D042783:Endothelial Cells / Q000378:metabolism*; D054324:Enterohemorrhagic Escherichia coli; D004797:Enzyme-Linked Immunosorbent Assay; D004927:Escherichia coli Infections / Q000150:complications / Q000378:metabolism; D005434:Flow Cytometry; D006463:Hemolytic-Uremic Syndrome / Q000378:metabolism / Q000382:microbiology; D006801:Humans; D007678:Kidney Glomerulus / Q000378:metabolism*; D007687:Kidney Tubules, Proximal / Q000378:metabolism*; D018613:Microscopy, Confocal; D020133:Reverse Transcriptase Polymerase Chain Reaction; D022641:Shiga Toxin 2 / Q000378:metabolism*
chemical_list: D018957:CD59 Antigens; D022641:Shiga Toxin 2; C051590:CD59 protein, human