An in vitro model for toxin-mediated vascular leak syndrome: ricin toxin A chain increases the permeability of human endothelial cell monolayers.
Vascular leak syndrome (VLS) is the dose-limiting toxicity observed in clinical trials of immunotoxins containing ricin toxin A chain (RTA). RTA itself is thought to cause VLS by damaging vascular endothelial cells, but the exact mechanism remains unclear. This is partially due to the paucity of appropriate models. To study VLS, we developed an in vitro model in which human umbilical vein-derived endothelial cells were first grown to confluence on microporous supports and then cultured under low pressure in the presence or absence of RTA. Endothelial cell barrier function was assessed by measuring the volume of fluid that passed through each monolayer per unit time. We found that RTA significantly increased monolayer permeability at times and concentrations consistent with the onset of VLS in patients treated with RTA-based immunotoxins. Scanning electron microscopy showed that intercellular gaps formed in endothelial monolayers exposed to RTA. Intercellular gap formation followed endothelial cell death caused by the enzymatic activity of RTA. We conclude that RTA is directly toxic to endothelial cells in vitro and speculate that this contributes to VLS in vivo.
Lindstrom
A L
AL
Department of Laboratory Medicine and Pathology, the Cancer Center, University of Minnesota, Minneapolis 55455, USA.
Erlandsen
S L
SL
Kersey
J H
JH
Pennell
C A
CA
D002199:Capillary Permeability / Q000187:drug effects*; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000187:drug effects*; D005353:Fibronectins / Q000494:pharmacology; D006801:Humans; D000922:Immunotoxins / Q000633:toxicity; D007365:Intercellular Junctions / Q000187:drug effects; D008855:Microscopy, Electron, Scanning; D008954:Models, Biological; D012276:Ricin / Q000493:pharmacokinetics / Q000633:toxicity*; D013917:Thrombin / Q000494:pharmacology; D013997:Time Factors
D005353:Fibronectins; D000922:Immunotoxins; D012276:Ricin; D013917:Thrombin