Flow cytometry detection of Shiga toxins in the blood from children with hemolytic uremic syndrome

Cytometry. Part B, Clinical Cytometry
Pier Luigi TazzariMaurizio Brigotti

Abstract

Hemolytic uremic syndrome (HUS) is the main cause of acute renal failure in early childhood. Most cases are due to intestinal infections from Escherichia coli strains (STEC) which produce by Shiga toxin (Stxs). Stx1 and Stx2 produced by STEC in the gut are absorbed into the circulation and, after binding on polymorphonuclear leukocytes (PMNs), are targeted to renal endothelium. The aim of the present work was the development of a method to detect Stxs bound on circulating PMNs and to diagnose STEC infections in patients with HUS. White blood cells isolated after erythrocytic lysis were incubated with anti-Stxs mouse monoclonal antibodies in the presence of human serum to saturate Fc receptors on PMNs. After incubation with fluorescein isothiocyanate-goat anti-mouse immunoglobulin G, flow cytometric analysis was used to demonstrate the cell-bound fluorescence. The method was quick (3 h), sensitive (femtomoles), and capable of detecting both Stxs. The presence of Stxs was detected on PMNs from six patients with HUS: four patients had serologic or microbiological evidence of STEC infection, whereas the other two patients had no evidence of STEC infection when employing the standard diagnostic methods. The method described is rapid...Continue Reading

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Citations

Sep 19, 2012·Nature Reviews. Nephrology·Howard TrachtmanRolf A K Stahl
Dec 18, 2010·The Pediatric Infectious Disease Journal·Maurizio BrigottiAlfredo Caprioli
Mar 22, 2006·Pediatric Research·Hélène DecaluweFrançois Proulx
Dec 4, 2012·Toxins·Chad L MayerDeborah J Stearns-Kurosawa
Apr 1, 2005·Current Opinion in Pediatrics·Richard Siegler, Robert Oakes
Apr 4, 2006·The Surgical Clinics of North America·Ai-Xuan L HoltermanRuth A Seeler
Oct 22, 2005·International Journal of Medical Microbiology : IJMM·Helge KarchMartina Bielaszewska
Oct 11, 2016·Journal of Internal Medicine·Diana KarpmanIda Arvidsson
Nov 22, 2017·Toxins·Annie VillyssonDiana Karpman
Jul 16, 2008·Journal of Leukocyte Biology·Maurizio BrigottiPier Luigi Tazzari
Dec 24, 2018·Cellular Microbiology·Maurizio BrigottiReinhard Würzner
Sep 3, 2010·The Biochemical Journal·Valentina ArfilliMaurizio Brigotti
Apr 8, 2010·Journal of Leukocyte Biology·Maurizio BrigottiAlfredo Caprioli
Sep 27, 2013·The Journal of Immunology : Official Journal of the American Association of Immunologists·Maurizio BrigottiMarco A Cassatella
Dec 24, 2015·The Journal of Immunology : Official Journal of the American Association of Immunologists·Domenica CarnicelliMaurizio Brigotti
Oct 30, 2007·Journal of Medical Microbiology·Thomas P GrienerGlen D Armstrong
Mar 3, 2021·The American Journal of Pathology·Elisa VarroneMaurizio Brigotti
Jun 25, 2015·Microbiology Spectrum·Diana Karpman, Anne-Lie Ståhl
Dec 23, 2014·Microbiology Spectrum·Angela R Melton-Celsa
Mar 23, 2005·Lancet·Phillip I TarrWayne L Chandler

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