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Thromb Haemost 2004; 92(05): 1066-1075
DOI: 10.1160/TH04-04-0241
Wound Healing and Inflammation/Infection
Schattauer GmbH

Neutrophils stimulated by apolipoprotein(a) generate fragments that are stronger inhibitors of plasmin formation than apo(a)

Leila B. Lamanuzzi
1   Department of Biochemistry & Molecular Biology & IBBE-CNR, University of Bari, Italy
3   INSERM U460, CHU Bichat-Claude Bernard, Paris, France
,
El Mostafa Mtairag
2   Hassan II University, School of Sciences I, Department of Biology, Casablanca, Morocco
3   INSERM U460, CHU Bichat-Claude Bernard, Paris, France
,
Gabriella Pepe
1   Department of Biochemistry & Molecular Biology & IBBE-CNR, University of Bari, Italy
,
Eduardo Anglés-Cano
3   INSERM U460, CHU Bichat-Claude Bernard, Paris, France
› Author AffiliationsFinancial support: Leila B. Lamanuzzi was a recipient of a Marie Curie Host Fellowship for Early Stage Training from the European Commission to Institut Fédératif de Recherche, Circulation Paris 7 (Director: Prof. Bernard Levy), and of an ESF (P.O.P. 1994-1999) fellowship. This work was supported in part by research grant Adrienne et Pierre Sommer from the Fondation de France.
Further Information

Publication History

Received 16 April 2004

Accepted after resubmission 16 August 2004

Publication Date:
04 December 2017 (online)

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Summary

Apolipoprotein(a), the plasminogen-like component of lipoprotein(a), is transformed into fragments by polymorphonuclear neutrophils (PMNs) elastase. Since stimulated PMNs express urokinase-type plasminogen activator (uPA), we sought to investigate the relevance of apo(a) fragmentation on plasminogen activation by neutrophils. Freshly isolated human PMNs stimulated by a 10 kringle recombinant apo(a), r-apo(a), activate plasminogen in a specific and saturable manner (Km = 476 ± 42 nM, Vmax = 896 ± 18 pmol min-1). This activation is prevented by amiloride, an inhibitor of u-PA, and ɛ-aminocaproic acid, ɛ-ACA, a lysine analogue that blocks plasminogen binding to PMNs. Stimulation of PMNs by apo(a) results in the formation of elastase-derived apo(a) fragments. These fragments produce a concentration-dependent decrease in the formation of plasmin. Addition of elastase inhibitors to PMNs prevented degradation of apo(a) and partially restored the formation of plasmin. In a similar manner, isolated r-apo(a) fragments were able to produce a 100% decrease in plasmin generation as compared to intact r-apo(a). These data indicate that apo(a) fragments produce a more pronounced inhibition in the generation of cellbound plasmin by uPA than the parent apo(a). These effects of apo(a) and its fragments were neutralised by a monoclonal antibody directed against the lysine-binding site of apo(a). This mechanism may be of biological relevance to the effects of Lp(a) in conditions where PMNs accumulate and release elastase, i.e. thrombus lysis and inflammatory lesions.

Keywords

Neutrophils - apolipoprotein(a) - elastase - plasminogen - proteolysis
 

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