Glycosphingolipid inhibition of the adhesion of thrombin-activated platelets to surfaces is potentiated by albumin.
Previous studies have shown that exogenous glycosphingolipids (GSLs) inhibit the adhesion of thrombin-activated platelets (TAP) to polystyrene plates coated with various RGD-ligands (where RGD is the peptide sequence Arg-Gly-Asp), suggesting that GSLs can modulate the platelet integrin receptor glycoprotein IIb-IIIa. However, albumin was always used as a plastic surface-blocking agent in these studies. In order to evaluate the role of albumin in these experiments, we studied the effect of various GSLs and albumin on the interaction between TAP and hydrophobic surfaces in a solid-phase assay using indium-111-labelled platelets and polystyrene plates. TAP (10(8) platelets/ml) adhered to polystyrene (half-saturation time 40 +/- 3 min) with a maximal adhesion density of 56 +/- 1 x 10(3) platelets/mm2. Platelet adhesion was only slightly affected (< 11% inhibition) by immobilized bovine serum albumin, immobilized mixed bovine brain gangliosides (MBG) or fluid-phase MBG. In contrast, fluid-phase MBG was an effective inhibitor of platelet adhesion to polystyrene (> 46% inhibition), but only after albumin was first immobilized to the plate. Covering albumin-coated polystyrene with MBG, followed by washing, was as effective as fluid-phase MBG at inhibiting platelet adhesion, thus indicating that a ganglioside-albumin interaction at the polystyrene surface was responsible for effective inhibition. When purified GSLs were substituted for MBG, it was found that all those tested (GT1b, GD1a, GM1, asialo GM1 and globoside) had similar inhibitory activity.
Department of Pathology, University of Iowa College of Medicine, Iowa City 52242.
M T
MT
Cunningham
J D
JD
Olson
T A
TA
Koerner
D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D002240:Carbohydrate Sequence; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D005732:Gangliosides / Q000494:pharmacology; D006028:Glycosphingolipids / Q000494:pharmacology*; D006801:Humans; D008969:Molecular Sequence Data; D015539:Platelet Activation / Q000187:drug effects; D011137:Polystyrenes / Q000378:metabolism*; D012709:Serum Albumin / Q000494:pharmacology*; D013917:Thrombin / Q000494:pharmacology
D005732:Gangliosides; D006028:Glycosphingolipids; D011137:Polystyrenes; D012709:Serum Albumin; D013917:Thrombin