A rapid and sensitive method to measure the functional activity of shiga toxins in human serum

Valentina Arfilli, Domenica Carnicelli, Gianluigi Ardissino, Erminio Torresani, Gaia Scavia, Maurizio Brigotti

Research output: Contribution to journalArticlepeer-review

Abstract

Shiga toxins (Stx) have a definite role in the development of hemolytic uremic syndrome in children with hemorrhagic colitis caused by pathogenic Stx-producing Escherichia coli (STEC) strains. The dramatic effects of these toxins on the microvasculature of different organs, particularly of the kidney, are well known, whereas there is no consensus on the mechanism by which Stx reach the endothelia of target organs and/or indirectly injure these body sites. We hereby describe a quick (4 h), radioactive, Raji cell-based method designed for the detection of Stx in human sera. The assay monitors the translation impairment induced by these powerful inhibitors of protein synthesis, which are identified properly by neutralizing their activity with specific monoclonal antibodies. By this method, we detected for the first time the functional activity of Stx in sera of STEC-infected patients during hemorrhagic colitis. Recent research has pointed to a dynamic process of Stx-induced renal intoxication in which concurrent and interactive steps are involved. Our rapid and specific method could be useful for studying the kinetics of Stx during the natural course of STEC infection and the interplay between Stx activity in serum and Stx presence in different blood fractions (neutrophils, monocytes, platelets, leukocyte-platelet aggregates, microvesicles, lipoproteins).

Original languageEnglish
Pages (from-to)4564-4576
Number of pages13
JournalToxins
Volume7
Issue number11
DOIs
Publication statusPublished - Nov 4 2015

Keywords

  • Eukaryotic protein synthesis
  • Hemolytic uremic syndrome
  • Raji cells
  • Shiga toxin-producing Escherichia coli

ASJC Scopus subject areas

  • Toxicology
  • Health, Toxicology and Mutagenesis

Fingerprint

Dive into the research topics of 'A rapid and sensitive method to measure the functional activity of shiga toxins in human serum'. Together they form a unique fingerprint.

Cite this