Biochemical characterisation of polycation-induced cytotoxicity to human vascular endothelial cells.
Pro-inflammatory effects of cationic proteins secreted by human granulocytes include induction of increased vascular permeability and oedema, which are likely to be mediated by damage to vascular endothelium. We have shown previously that a series of synthetic polycationic amino acids produce a dose-, time- and Mr-dependent inhibition of [3H]leucine or [3H]thymidine incorporation into macromolecules by human umbilical vein endothelial cells, and that the extent of inhibition was correlated with changes in cell morphology, with release of cytoplasmic constituents and was irreversible. The experiments reported here characterise further the requirements for the induction of cytotoxicity by polycations. We have found that the extent of inhibition is related to both the identity of the monomer, for polymers of Mr 40,000 the order is ornithine greater than lysine greater than arginine, and to its configuration; poly-D-lysines are more potent inhibitors than poly-L-lysines of similar Mr. Only brief exposure to the agonist is required, 90% inhibition occurred after 10 min of exposure to poly-L-lysine (Mr 90,000). Treatment of endothelial cells with neuraminidase, heparinase, hyaluronidase, chondroitinase or trypsin did not reduce their susceptibility to polylysine. Inhibition of microtubule or microfilament formation also had no effect on polylysine cytotoxicity, indicating that internalisation of the polymer was not a prerequisite for the effect. Inhibition of protein synthesis or pretreatment with simple sugars likewise failed to block the effects of polylysine treatment. Natural cationic proteins exerted similar effects on endothelial cells, the extent of the effect apparently being related to the pI of the protein.(ABSTRACT TRUNCATED AT 250 WORDS)
Morgan
D M
DM
Section of Vascular Biology, MRC Clinical Research Centre, Harrow, Middlesex, UK.
Larvin
V L
VL
Pearson
J D
JD
D000596:Amino Acids / Q000494:pharmacology; D001665:Binding Sites; D002412:Cations; D002462:Cell Membrane / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007930:Leucine / Q000378:metabolism; D008970:Molecular Weight; D010455:Peptides / Q000494:pharmacology; D011073:Polyamines; D000071228:Polyelectrolytes; D011107:Polylysine / Q000494:pharmacology; D011108:Polymers / Q000378:metabolism / Q000494:pharmacology*; D013936:Thymidine / Q000378:metabolism
D000596:Amino Acids; D002412:Cations; D007930:Leucine; D010455:Peptides; D011073:Polyamines; D000071228:Polyelectrolytes; D011107:Polylysine; D011108:Polymers; D013936:Thymidine