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Thrombin-activated human platelets release two NAP-2 variants that stimulate polymorphonuclear leukocytes

P Piccardoni1, V Evangelista, A Piccoli

  • 1Istituto di Ricerche Farmacologiche, Mario Negri, Consorzio Mario Negri Sud, Santa Maria Imbaro, Italy. Piccardoni@cmns.mnegri.it

Thrombosis and Haemostasis
|November 1, 1996
PubMed
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Thrombin-activated platelets release specific neutrophil-activating peptide 2 (NAP-2) variants that directly stimulate polymorphonuclear leukocytes (PMN). These active variants are released without further proteolytic processing, clarifying platelet-PMN interactions in inflammation and thrombosis.

Area of Science:

  • Hematology
  • Immunology
  • Biochemistry

Background:

  • Activated human platelets release proteins that increase intracellular calcium in polymorphonuclear leukocytes (PMN).
  • The specific platelet-derived factors responsible for PMN stimulation were not fully characterized.

Purpose of the Study:

  • To identify and characterize the platelet-released products that stimulate PMN.
  • To elucidate the mechanism of generation and release of these PMN-stimulating factors.

Main Methods:

  • Isolation of PMN-stimulating activity from platelet supernatant using Fast Protein Liquid Chromatography (FPLC) and High-Performance Liquid Chromatography (HPLC).
  • N-terminal sequence analysis of purified peptides.
  • Heparin-affinity chromatography to assess heparin-binding properties.

Related Experiment Videos

  • Cross-desensitization experiments and antibody inhibition assays.
  • Assessment of protease inhibitor effects on PMN-stimulating activity.

    • Main Results:

    • Purified fractions consisted mainly of truncated forms of connective tissue-activating peptide III (CTAP-III), identified as neutrophil-activating peptide 2 (NAP-2) variants.
    • These NAP-2 variants are heparin-binding proteins and account for ATP-independent PMN stimulation.
    • NAP-2 variants were released in an active form and not generated by extracellular proteolytic cleavage from platelet or leukocyte proteases.

    Conclusions:

    • Thrombin-activated platelets release active NAP-2 variants that directly stimulate PMN.
    • The release mechanism does not involve extracellular proteolytic processing.
    • This finding advances the understanding of platelet-PMN crosstalk in thrombosis and inflammation.