title: Tumor necrosis factor-alpha enhances platelet activation via cathepsin G released from neutrophils.
abstract: In this paper we show that TNF-alpha enhances platelet activation. Experiments were performed on a human polymorphonuclear neutrophil (PMN)-platelet cooperation system in which PMN, stimulated by FMLP, release cathepsin G (Cat.G), a serine proteinase responsible for the activation of nearby platelets. Pretreatment of the mixed cell suspension with 5 ng/ml TNF-alpha resulted in a strong platelet activation (37.7 +/- 3.2% aggregation; 46.0 +/- 14.4% serotonin release) in response to a weak concentration of FMLP (1.25 x 10(-8) M) inducing by itself only 7.7 +/- 4.0% of aggregation and 3.8 +/- 4.1% of serotonin release (mean +/- SD; n = 10). This effect was concentration dependent (maximum between 5 and 10 ng/ml) and was optimal for a brief preincubation time (5 min). Under these experimental conditions the target of TNF-alpha was PMN, as shown by beta-glucuronidase release. The observed potentiation was modified neither by 0.1 mM acetyl salicylic acid (a cyclo-oxygenase inhibitor) nor by 0.1 mM BN 52021 (a platelet-activating factor antagonist), while such a phenomenon was fully inhibited by 20 micrograms/ml eglin C, a strong and specific inhibitor of the human granulocytic proteinases, elastase and Cat.G. In fact, full inhibition was also observed with 300 nM alpha-1-antichymotrypsin, a specific inhibitor of Cat.G. This clear-cut evidence of Cat.G involvement was substantiated by the enhancement of Cat.G release from FMLP-activated PMN primed with TNF-alpha. These results demonstrate that the priming of PMN by TNF-alpha may modulate the activation of other inflammatory cells, particularly of platelets. It is hypothesized that this phenomenon could contribute to pulmonary pathologies, and more specifically to the adult respiratory distress syndrome, a disease for which PMN, platelet and TNF-alpha involvement has been proposed.
authors.affiliation: Unité de Pharmacologie Cellulaire, Unité Associée IP/INSERM 285, Institut Pasteur, Paris, France.
authors.forename: P
authors.initials: P
authors.lastname: Renesto
authors.forename: M
authors.initials: M
authors.lastname: Chignard
mesh_terms: D056649:Cathepsin G; D002403:Cathepsins / Q000502:physiology*; D005966:Glucuronidase / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000502:physiology*; D015539:Platelet Activation / Q000187:drug effects*; D011506:Proteins; D012697:Serine Endopeptidases; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D012701:Serotonin / Q000378:metabolism; D015843:Serpins; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D000514:alpha 1-Antichymotrypsin / Q000494:pharmacology
chemical_list: D011506:Proteins; D015842:Serine Proteinase Inhibitors; D015843:Serpins; D014409:Tumor Necrosis Factor-alpha; D000514:alpha 1-Antichymotrypsin; C028656:eglin proteinase inhibitors; D012701:Serotonin; D009240:N-Formylmethionine Leucyl-Phenylalanine; D005966:Glucuronidase; D002403:Cathepsins; D012697:Serine Endopeptidases; C534094:CTSG protein, human; D056649:Cathepsin G