title: Neutrophil-derived proteinase 3 induces kallikrein-independent release of a novel vasoactive kinin.
abstract: The kinin-forming pathway is activated on endothelial cells and neutrophils when high-molecular weight kininogen (HK) is cleaved by plasma kallikrein liberating bradykinin, a potent mediator of inflammation. Kinins are released during inflammatory conditions such as vasculitis, associated with neutrophil influx around blood vessels. Some patients with vasculitis have elevated plasma levels of neutrophil-derived proteinase 3 (PR3) and anti-PR3 Abs. This study investigated if neutrophil-derived PR3 could induce activation of the kinin pathway. PR3 incubated with HK, or a synthetic peptide derived from HK, induced breakdown and release of a novel tridecapeptide termed PR3-kinin, NH(2)-MKRPPGFSPFRSS-COOH, consisting of bradykinin with two additional amino acids on each terminus. The reaction was specific and inhibited by anti-PR3 and alpha(1)-antitrypsin. Recombinant wild-type PR3 incubated with HK induced HK breakdown, whereas mutated PR3, lacking enzymatic activity, did not. PR3-kinin bound to and activated human kinin B(1) receptors, but did not bind to B(2) receptors, expressed by transfected HEK293 cells in vitro. In human plasma PR3-kinin was further processed to the B(2) receptor agonist bradykinin. PR3-kinin exerted a hypotensive effect in vivo through both B(1) and B(2) receptors as demonstrated using wild-type and B(1) overexpressing rats as well as wild-type and B(2) receptor knockout mice. Neutrophil extracts from vasculitis patients and healthy controls contained comparable amounts of PR3 and induced HK proteolysis, an effect that was abolished when PR3 was immunoadsorbed. Neutrophil-derived PR3 can proteolyze HK and liberate PR3-kinin, thereby initiating kallikrein-independent activation of the kinin pathway.
authors.affiliation: Department of Pediatrics, Clinical Sciences Lund, Lund University, Lund, Sweden.
authors.forename: Robin
authors.initials: R
authors.lastname: Kahn
authors.forename: Thomas
authors.initials: T
authors.lastname: Hellmark
authors.forename: L M Fredrik
authors.initials: LM
authors.lastname: Leeb-Lundberg
authors.forename: Nasrin
authors.initials: N
authors.lastname: Akbari
authors.forename: Mihail
authors.initials: M
authors.lastname: Todiras
authors.forename: Tor
authors.initials: T
authors.lastname: Olofsson
authors.forename: Jörgen
authors.initials: J
authors.lastname: Wieslander
authors.forename: Anders
authors.initials: A
authors.lastname: Christensson
authors.forename: Kerstin
authors.initials: K
authors.lastname: Westman
authors.forename: Michael
authors.initials: M
authors.lastname: Bader
authors.forename: Werner
authors.initials: W
authors.lastname: Müller-Esterl
authors.forename: Diana
authors.initials: D
authors.lastname: Karpman
mesh_terms: D000293:Adolescent; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D001920:Bradykinin / Q000097:blood; D002648:Child; D006801:Humans; D007610:Kallikreins / Q000378:metabolism; D007704:Kininogens / Q000378:metabolism; D007705:Kinins / Q000378:metabolism*; D008297:Male; D051379:Mice; D008875:Middle Aged; D053146:Myeloblastin / Q000235:genetics / Q000378:metabolism*; D009504:Neutrophils / Q000201:enzymology*; D011485:Protein Binding; D051381:Rats
chemical_list: D007704:Kininogens; D007705:Kinins; D007610:Kallikreins; D053146:Myeloblastin; D001920:Bradykinin