Platelet activation by fMLP-stimulated polymorphonuclear leukocytes: The activity of cathepsin G is not prevented by antiproteinases

Virgilio Evangelista, Grazyna Rajtar, Giovanni De Gaetano, James G. White, Chiara Cerletti

Research output: Contribution to journalArticle

Abstract

Human polytnorphonuclear leukocytes (PMN) activated by fMLP (in the presence of CaCl2, fibrinogen, and cytochalasin B) were able to induce aggregation, cytoplasmic Ca2+ increase and thromboxane A2 production in coincubated autologousplatelets. Cell-freesupernatantspreparedfromn-formyl-methionyl-leucyl-phenylalanine (fMLP)-stimulated PMN were ble also to induce platelet activation. Antibodies against cathepsin G and different serin protease inhibitors completely suppressed the activity of PMN-derived supernatants, indicating that cathepsin G is the major platelet activator released by PMN in our system. However, antiproteinases only partially affected platelet activation induced by PMN in mixed cell suspensions. Superoxide dismutase and catalase added to the cell suspension did not affect platelet activation nor potentiated serin protease inhibitors, making a role for short-lived oxygen radicals in our experimental system unlikely. Electron microscopic observation of stirred mixed cell suspensions preincubated for 2 minutes at 37° C before stimulation showed a close PMN-platelets contact without any morphologic or biochemical event suggesting platelet activation. Preincubation of the cells without stirring to minimize PMN-platelet interaction before stimulation did not modify subsequent aggregation and platelet cytoplasmic Ca2+ increase in control samples. However, in this condition trypsin inhibitor from soybean completely prevented PMM-induced platelet activation. In samples preincubated without stirring in the presence of the antiproteinase, activated PMN sticked together but platelets preserved their discoid shape and did not appear significantly activated. We propose that membrane-to-membrane contact could create a microenvironment in which cathepsin G, discharged from stimulated PMN on adherent platelets, is protected from antiproteinases.

Original languageEnglish
Pages (from-to)2379-2388
Number of pages10
JournalBlood
Volume77
Issue number11
Publication statusPublished - Jun 1 1991

ASJC Scopus subject areas

  • Hematology

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