Bacterial exotoxins and endothelial permeability for water and albumin in vitro.

Effects of Staphylococcus aureus alpha-toxin and Pseudomonas aeruginosa cytotoxin on the permeability of an endothelial monolayer were studied. Porcine pulmonary artery endothelial cells were grown on a polycarbonate membrane, mounted in a chamber, and exposed to a continuous hydrostatic pressure of 10 cmH2O. On application of this trans-endothelial pressure, endothelial monolayer became "sealed," i.e., the filtration rate for water decreased and the reflection coefficient for albumin increased, reaching a plateau after 1-2 h. Sealed monolayer had a hydraulic conductivity of 2.1 X 10(-6) cm.s-1.cmH2O and an albumin reflection coefficient of 0.73. Permeability of the monolayer was increased on addition of an excess of EDTA and reversed on readdition of calcium. Within 60-90 min after addition of 1 microgram/ml alpha-toxin, the filtration rate increased 75-fold, and the albumin reflection coefficient dropped to 0.20. These changes in permeability were accompanied by cell retraction and formation of large intercellular gaps between endothelial cells. Effects of alpha-toxin were abolished by preincubation with neutralizing antibodies and by inhibitors of calmodulin function. Pseudomonas aeruginosa cytotoxin (25 and 50 micrograms/ml) also increased the permeability of the endothelial monolayer, but it was only about one-third as effective as alpha-toxin.

[1]  H B Hechtman,et al.  Serotonin, norepinephrine, and histamine mediation of endothelial cell barrier function in vitro , 1986, Journal of cellular physiology.

[2]  M. Baetscher,et al.  An in vitro system for measuring endothelial permeability under hydrostatic pressure. , 1983, Experimental cell research.

[3]  F. Lutz.  Purification of a cytotoxic protein from Pseudomonas aeruginosa. , 1979, Toxicon : official journal of the International Society on Toxinology.

[4]  H. Ochs,et al.  Activated neutrophils disrupt endothelial monolayer integrity by an oxygen radical-independent mechanism. , 1985, Laboratory investigation; a journal of technical methods and pathology.

[5]  T. Pollard,et al.  Actin filament stress fibers in vascular endothelial cells in vivo. , 1983, Science.

[6]  B. Rippe,et al.  Effects of histamine, serotonin, and norepinephrine on circulation of dog lungs. , 1984, Journal of applied physiology: respiratory, environmental and exercise physiology.

[7]  R. Rogers,et al.  Adult respiratory-distress syndrome: changing concepts of lung injury and repair. , 1982, The New England journal of medicine.

[8]  S. Bhakdi,et al.  Damage to mammalian cells by proteins that form transmembrane pores. , 1987, Reviews of physiology, biochemistry and pharmacology.

[9]  W. Seeger,et al.  Mechanism of leukotriene generation in polymorphonuclear leukocytes by staphylococcal alpha-toxin , 1987, Infection and immunity.

[10]  D. Drenckhahn,et al.  Stress fibers in the splenic sinus endothelium in situ: molecular structure, relationship to the extracellular matrix, and contractility , 1986, The Journal of cell biology.

[11]  W. Seeger,et al.  Staphylococcal alpha-toxin elicits hypertension in isolated rabbit lungs. Evidence for thromboxane formation and the role of extracellular calcium. , 1984, The Journal of clinical investigation.

[12]  B. Rippe,et al.  Effects of plasma- and cell-free perfusates on filtration coefficient of perfused canine lungs. , 1985, Journal of applied physiology.

[13]  S. Bhakdi,et al.  On the mechanism of membrane damage by Staphylococcus aureus alpha- toxin , 1981, The Journal of cell biology.

[14]  N. Suttorp,et al.  Lung cell oxidant injury. Enhancement of polymorphonuclear leukocyte-mediated cytotoxicity in lung cells exposed to sustained in vitro hyperoxia. , 1982, The Journal of clinical investigation.

[15]  M. Peach,et al.  Role of Endothelial Cell Cytoskeleton in Control of Endothelial Permeability , 1982, Circulation research.

[16]  S Bhakdi,et al.  Staphylococcal alpha-toxin: oligomerization of hydrophilic monomers to form amphiphilic hexamers induced through contact with deoxycholate detergent micelles. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[17]  D. Shasby,et al.  Effects of calcium on transendothelial albumin transfer and electrical resistance. , 1986, Journal of applied physiology.

[18]  M Cereijido,et al.  Experimental modulation of occluding junctions in a cultured transporting epithelium , 1980, The Journal of cell biology.

[19]  K. Failing,et al.  Cytotoxic protein from Pseudomonas aeruginosa: formation of hydrophilic pores in Ehrlich ascites tumor cells and effect on cell viability. , 1987, Toxicon : official journal of the International Society on Toxinology.

[20]  R. Franke,et al.  Induction of human vascular endothelial stress fibres by fluid shear stress , 1984, Nature.

[21]  R. Bizios,et al.  Thrombin‐induced increase in albumin permeability across the endothelium , 1986, Journal of cellular physiology.

[22]  W. Seeger,et al.  Pseudomonas aeruginosa cytotoxin stimulates prostacyclin production in cultured pulmonary artery endothelial cells: Membrane attack and calcium influx , 1985, Journal of cellular physiology.

[23]  W. Seeger,et al.  Pulmonary microvascular injury induced by Pseudomonas aeruginosa cytotoxin in isolated rabbit lungs , 1986, Infection and Immunity.