Examination of the toxicity of several protein toxins of plant origin using bovine pulmonary endothelial cells.
The bovine pulmonary endothelial (BPE) cell line was examined as a model to study the toxicity of ricin and abrin toxins currently under investigation. The BPE cell line was examined because ricin has been shown to bind to endothelial cells. Cell viability was assessed using several different biochemical parameters including growth (DNA by binding of gentian violet stain), mitochondrial function (succinate dehydrogenase activity) using MTT and lysosomal integrity (neutral red retention assay). In order to compare toxicities and investigate potential protective compounds, concentrations of toxins causing death of 50% and 70% of the (control) cell population (LC50 and LC70, respectively) were determined. It is concluded that while ricin and abrin share a common mechanism of action ricin is slightly less toxic than abrin. BPE cells are a good model for future mechanistic studies and particularly for initial phase screening of potentially therapeutic compounds. Carbohydrates were used in an attempt to examine which receptor types were involved in the binding and uptake of ricin and abrin by the cell line. It was found that only high concentrations of galactose prevented lethality while mannose apparently had no effect. Furthermore, the molar excess of carbohydrate to toxin required in order to achieve protection indicated that this would be an impractical approach to adopt in vivo.
Chemical and Biological Defence Establishment, Porton Down, Salisbury, Wiltshire, UK.
G D
GD
Griffiths
C D
CD
Lindsay
D G
DG
Upshall
D000036:Abrin / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000633:toxicity*; D000818:Animals; D002417:Cattle; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004727:Endothelium / Q000187:drug effects; D005690:Galactose / Q000494:pharmacology; D005840:Gentian Violet; D007928:Lethal Dose 50; D008168:Lung / Q000187:drug effects*; D008351:Mannans / Q000494:pharmacology; D008358:Mannose / Q000494:pharmacology; D009499:Neutral Red; D012276:Ricin / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000633:toxicity*; D013997:Time Factors
D008351:Mannans; D000036:Abrin; D009499:Neutral Red; D012276:Ricin; D005840:Gentian Violet; D008358:Mannose; D005690:Galactose