title: Association of Shiga toxin glycosphingolipid receptors with membrane microdomains of toxin-sensitive lymphoid and myeloid cells.
abstract: Glycosphingolipids (GSLs) of the globo-series constitute specific receptors for Shiga toxins (Stxs) released by certain types of pathogenic Escherichia coli strains. Stx-loaded leukocytes may act as transporter cells in the blood and transfer the toxin to endothelial target cells. Therefore, we performed a thorough investigation on the expression of globo-series GSLs in serum-free cultivated Raji and Jurkat cells, representing B- and T-lymphocyte descendants, respectively, as well as THP-1 and HL-60 cells of the monocyte and granulocyte lineage, respectively. The presence of Stx-receptors in GSL preparations of Raji and THP-1 cells and the absence in Jurkat and HL-60 cells revealed high compliance of solid-phase immunodetection assays with the expression profiles of receptor-related glycosyltransferases, performed by qRT-PCR analysis, and Stx2-caused cellular damage. Canonical microdomain association of Stx GSL receptors, sphingomyelin, and cholesterol in membranes of Raji and THP-1 cells was assessed by comparative analysis of detergent-resistant membrane (DRM) and nonDRM fractions obtained by density gradient centrifugation and showed high correlation based on nonparametric statistical analysis. Our comprehensive study on the expression of Stx-receptors and their subcellular distribution provides the basis for exploring the functional role of lipid raft-associated Stx-receptors in cells of leukocyte origin.
authors.affiliation: Institute for Hygiene, University of Münster, D-48149 Münster, Germany.
authors.forename: Ivan U
authors.initials: IU
authors.lastname: Kouzel
authors.affiliation: Institute for Hygiene, University of Münster, D-48149 Münster, Germany.
authors.forename: Gottfried
authors.initials: G
authors.lastname: Pohlentz
authors.affiliation: Institute for Hygiene, University of Münster, D-48149 Münster, Germany.
authors.forename: Wiebke
authors.initials: W
authors.lastname: Storck
authors.affiliation: Institute for Hygiene, University of Münster, D-48149 Münster, Germany.
authors.forename: Lena
authors.initials: L
authors.lastname: Radamm
authors.affiliation: Institute for Hygiene, University of Münster, D-48149 Münster, Germany.
authors.forename: Petra
authors.initials: P
authors.lastname: Hoffmann
authors.affiliation: Institute for Hygiene, University of Münster, D-48149 Münster, Germany.
authors.forename: Martina
authors.initials: M
authors.lastname: Bielaszewska
authors.affiliation: Institute for Hygiene, University of Münster, D-48149 Münster, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Bauwens
authors.affiliation: Institute of Infectiology, University of Münster, D-48149 Münster, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Cichon
authors.affiliation: Institute of Infectiology, University of Münster, D-48149 Münster, Germany.
authors.forename: M Alexander
authors.initials: MA
authors.lastname: Schmidt
authors.affiliation: Institute for Hygiene, University of Münster, D-48149 Münster, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Mormann
authors.affiliation: Institute for Hygiene, University of Münster, D-48149 Münster, Germany.
authors.forename: Helge
authors.initials: H
authors.lastname: Karch
authors.affiliation: Institute for Hygiene, University of Münster, D-48149 Münster, Germany; Interdisciplinary Center for Clinical Research (IZKF), University of Münster, D-48149 Münster, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Müthing
mesh_terms: D015153:Blotting, Western; D002460:Cell Line; D049109:Cell Proliferation; D004591:Electrophoresis, Polyacrylamide Gel; D005700:Galactosyltransferases / Q000378:metabolism; D005915:Globosides / Q000378:metabolism; D006028:Glycosphingolipids / Q000378:metabolism*; D018922:HL-60 Cells; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism*; D021962:Membrane Microdomains / Q000378:metabolism*; D022423:Myeloid Cells / Q000378:metabolism*; D017350:N-Acetylgalactosaminyltransferases / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D022621:Shiga Toxin / Q000378:metabolism*; D014281:Trihexosylceramides / Q000378:metabolism
chemical_list: D005915:Globosides; D006028:Glycosphingolipids; D014281:Trihexosylceramides; C024032:globotetraosylceramide; C018549:globotriaosylceramide; D022621:Shiga Toxin; D005700:Galactosyltransferases; D017350:N-Acetylgalactosaminyltransferases; C045097:UDP-galactose-lactosylceramide alpha 1-4-galactosyltransferase; C023220:(N-acetylneuraminyl)-galactosylglucosylceramide N-acetylgalactosaminyltransferase