title: Human endothelial cell activation and mediator release in response to the bacterial exotoxins Escherichia coli hemolysin and staphylococcal alpha-toxin.
abstract: Escherichia coli hemolysin (HlyA) and Staphylococcus aureus alpha-toxin are membrane-perturbating bacterial exotoxins that have been implicated as significant virulence factors in human diseases. We investigated the capacity of these toxins to cause cell activation and mediator release in human endothelial cells, compared with the efficacies of thrombin and the Ca2+ ionophore A23187. Concentration ranges tested were 1 to 1000 ng/ml (HlyA), 0.01 to 10 micro/ml (alpha-toxin), 0.01 to 10 U/ml (thrombin), and 0.01 to 10 microM (A23187). All stimuli caused dose-dependent generation of platelet-activating factor, nitric oxide, and prostaglandin I2. HlyA and thrombin effected time- and dose-dependent accumulation of large quantities of inositol phosphates, with maximum effects at 100 ng/ml and 1 U/ml, respectively. Corresponding time course and dose dependency were noted for HlyA-elicited diacylglycerol formation. In contrast, only the highest concentrations of alpha-toxin (10 microg/ml) and A23187 (10 microM) effected some moderate inositol phosphate accumulation, and this was suppressed in the presence of the platelet-activating factor antagonist WEB 2086. Metabolic and secretory responses elicited by alpha-toxin were dependent on the presence of extracellular Ca2+. We conclude that both HlyA and alpha-toxin are potent inductors of inflammatory and vasodilatory mediators in human endothelial cells. HlyA-elicited effects may proceed predominantly via activation of the phosphatidylinositol hydrolysis-related signal transduction pathway, whereas transmembrane Ca2+ flux appears to be the major event underlying the release of mediators in response to alpha-toxin. These toxin properties may contribute to vasoregulatory and inflammatory disturbances encountered in states of severe infection and sepsis.
authors.affiliation: Department of Internal Medicine, Justus-Liebig University, Giessen, Germany.
authors.forename: F
authors.initials: F
authors.lastname: Grimminger
authors.forename: F
authors.initials: F
authors.lastname: Rose
authors.forename: U
authors.initials: U
authors.lastname: Sibelius
authors.forename: M
authors.initials: M
authors.lastname: Meinhardt
authors.forename: B
authors.initials: B
authors.lastname: Pötzsch
authors.forename: R
authors.initials: R
authors.lastname: Spriestersbach
authors.forename: S
authors.initials: S
authors.lastname: Bhakdi
authors.forename: N
authors.initials: N
authors.lastname: Suttorp
authors.forename: W
authors.initials: W
authors.lastname: Seeger
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000633:toxicity*; D001427:Bacterial Toxins / Q000633:toxicity*; D002118:Calcium / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D011464:Epoprostenol / Q000096:biosynthesis; D029968:Escherichia coli Proteins; D006460:Hemolysin Proteins / Q000633:toxicity*; D006801:Humans; D009569:Nitric Oxide / Q000096:biosynthesis; D010716:Phosphatidylinositols / Q000378:metabolism; D010972:Platelet Activating Factor / Q000096:biosynthesis; D051381:Rats; D013917:Thrombin / Q000494:pharmacology
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; D029968:Escherichia coli Proteins; D006460:Hemolysin Proteins; C036507:Hlya protein, E coli; D010716:Phosphatidylinositols; D010972:Platelet Activating Factor; C021091:staphylococcal alpha-toxin; D009569:Nitric Oxide; D011464:Epoprostenol; D013917:Thrombin; D002118:Calcium