Effects of TNFα on verocytotoxin cytotoxicity in purified human glomerular microvascular endothelial cells
| Autor: | Victor W.M. van Hinsbergh, L.P.W.J. van den Heuvel, T. J. A. N. Van Der Velden, P.A. van Setten, Karel J.M. Assmann, N.C.A.J. van de Kar, L.A.H. Monnens, J.D. Mahan, Mario A. Vermeer |
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| Rok vydání: | 1997 |
| Předmět: |
medicine.medical_specialty
Immunologische ontstekingsprocessen in de nier Endothelium Cell Survival medicine.medical_treatment Bacterial Toxins Kidney Glomerulus Receptors Cell Surface Cell Separation Biology Shiga Toxin 1 Kidney Shiga Toxin 2 Internal medicine Inflammatory reactions in the kidneys medicine Humans Cytotoxicity GeneralLiterature_REFERENCE(e.g. dictionaries encyclopedias glossaries) Cells Cultured Tumor Necrosis Factor-alpha Immunity Brain Epidemiologie en pathogenese van het hemolytisch uremisch syndroom Molecular biology In vitro Epidemiology and pathogenesis of the hemolytic uremic syndrome Endothelial stem cell Endocrinology medicine.anatomical_structure Cytokine Cell culture Nephrology Cell Aging Protein Biosynthesis Hemolytic-Uremic Syndrome Tumor necrosis factor alpha Endothelium Vascular Cell aging |
| Zdroj: | Kidney International. Supplement, 51, pp. 1245-1256 Kidney International. Supplement, 51, 4, pp. 1245-1256 Kidney International. Supplement, 51, 1245-1256 |
| ISSN: | 0085-2538 2157-1724 |
| Popis: | Effects of TNFα and verocytotoxin cytotoxicity in human glomerular microvascular endothelial cells. In the pathogenesis of the hemolytic uremic syndrome (HUS), endothelial damage of glomeruli and arterioles of the kidney appears to play a central role. Previous studies have shown that verocytotoxin-1 (VT-1) cytotoxicity on human vein endothelial cells require additional stimuli, in particular the inflammatory mediator tumor necrosis factor alpha (TNFα). In this study the effects of VT on human glomerular microvascular endothelial cells (GMVEC) were examined. A reproducible method was developed for the isolation and purification of large numbers of highly purified GMVEC. The obtained GMVEC were over 99% pure; their endothelial origin was demonstrated by the expression of the endothelial antigens von Willebrand factor, EN-4, PECAM-1 and V,E-cadherin. Upon stimulation with TNFα the cells expressed the endothelial-specific adhesion molecule E-selectin. A limited number of fenestral structures was observed by scanning electron microscopy (SEM), suggesting glomerular origin of the endothelial cells. Cytotoxicity of VT-1 to GMVEC was evaluated by determination of the number of viable adherent cells and by assay of overall protein synthesis after exposure to varying concentrations of VT-1. In non-stimulated GMVEC, cytotoxicity of VT-1 was inversely related to the degree and duration of confluence, subconfluent cells being the most sensitive. In highly confluent GMVEC, VT cytotoxicity required pre-exposure of the cells to the inflammatory mediator TNFα, which induced an increase in the number of VT receptors on GMVEC. Thin layer chromatography of extracted glycolipids from the GMVEC showed binding of VT-1 to globotriaosylceramide (Gb3), known to be the functional receptor for VT. There were no major differences in protein synthesis inhibition with equal concentrations VT-1 and VT-2. In conclusion, in this study we provide a reproducible method to isolate, purify and culture well characterized human GMVEC on a routine basis. In vitro studies with these GMVEC demonstrate that VT cytotoxicity depends on the degree of confluence and the additional preexposure to the inflammatory mediator TNFα. These observations provide further insight into the complex events that may occur in glomeruli in the pathogenesis of HUS. |
| Databáze: | OpenAIRE |
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