Inhibition of leukocyte and platelet aggregation in vitro by antithrombin

Donatella Gritti, Chiara Gasparetto, Angela Malinverno, G. Ricevuti

Research output: Contribution to journalArticlepeer-review

Abstract

Glycosaminoglycan-mediated aggregation of cells occurs through adhesion mechanisms in which heparan sulfate chains bind to counter receptors on these cells. As antithrombin interacts with heparan sulfate proteoglycans through its heparin-binding domain and inhibits leukocyte adhesion in ischaemia/reperfusion, it may affect leukocyte aggregation. Leukocyte aggregation was therefore monitored in vitro as the increase in transmission of light through stirred suspensions in a platelet aggregometer. Aggregation curves were quantified as the area under the curve in the first 6 min following stimulation. Leukocytes in platelet-rich plasma were obtained from heparinized whole blood of healthy donors by centrifugation; the ratio of leukocytes to platelets was about 1:45, and the final concentration of autologous plasma was 80%. Neutrophils were purified by dextran sedimentation, density centrifugation, and hypotonic lysis of erythrocytes. Aggregation was induced by phytohaemagglutinin (0.24 mg/mL) or formyl-Met-Leu-Phe (0.2 × 10-6 M), with or without various concentrations of antithrombin. During the observation period (6 min) no aggregation of leukocytes in platelet-rich plasma or isolated neutrophils could be induced either with medium or with antithrombin (0.2 × 100 to 0.2 × 10-6 IU/mL). Addition of phytohaemagglutinin or formyl-Met-Leu-Phe stimulated aggregation of leukocytes in platelet-rich plasma and neutrophils to different extents. Additional presence of antithrombin significantly inhibited phytohaemagglutinin-induced aggregation of leukocytes in platelet-rich plasma, whereas formyl-Met-Leu-Phe-induced aggregation was not affected by antithrombin. Data show that in the presence of plasma and platelets, aggregation of normal white blood cells after stimulation with lectin but not with chemotaxin is inhibited by antithrombin, suggesting specific interactions of antithrombin with lectin-activated processes of neutrophil aggregation that occur in the presence of platelets and/or plasma.

Original languageEnglish
Pages (from-to)97-99
Number of pages3
JournalActa Medica Austriaca
Volume29
Issue number3
DOIs
Publication statusPublished - 2002

Keywords

  • Aggregation
  • Antithrombin
  • Inflammation
  • Leukocytes
  • Plasma
  • Platelets

ASJC Scopus subject areas

  • Medicine(all)

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