title: Regulation of leukocyte recruitment by polypeptides derived from high molecular weight kininogen.
abstract: Proteolytic cleavage of single-chain, high molecular weight kininogen (HK) by kallikrein releases the short-lived vasodilator bradykinin and leaves behind a two-chain, high molecular weight kininogen (HKa) reported to bind to the beta2-integrin Mac-1 (CR3, CD11b/CD18, alphaMbeta2) on neutrophils and exert antiadhesive properties by binding to the urokinase receptor (uPAR) and vitronectin. We define the molecular mechanisms for the antiadhesive effects of HK related to disruption of beta2-integrin-mediated cellular interactions in vitro and in vivo. In a purified system, HK and HKa inhibited the binding of soluble fibrinogen and ICAM-1 to immobilized Mac-1, but not the binding of ICAM-1 to immobilized LFA-1 (CD11a/CD18, alphaLbeta2). This inhibitory effect could be attributed to HK domain 5 and to a lesser degree to HK domain 3, consistent with the requirement of both domains for binding to Mac-1. Accordingly, HK, HKa, and domain 5 inhibited the adhesion of Mac-1 but not LFA-1-transfected K562 human erythroleukemic cells to ICAM-1. Moreover, adhesion of human monocytic cells to fibrinogen and to human endothelial cells was blocked by HK, HKa, and domain 5. By using peptides derived from HK domain 5, the sequences including amino acids H475-G497 (and to a lesser extent, G440-H455) were identified as responsible for the antiadhesive effect, which was independent of uPAR. Finally, administration of domain 5 into mice, followed by induction of thioglycollate-provoked peritonitis, decreased the recruitment of neutrophils by approximately 70% in this model of acute inflammation. Taken together, HKa (and particularly domain 5) specifically interacts with Mac-1 but not with LFA-1, thereby blocking Mac-1-dependent leukocyte adhesion to fibrinogen and endothelial cells in vitro and in vivo and serving as a novel endogenous regulator of leukocyte recruitment into the inflamed tissue.
authors.affiliation: Institute for Biochemistry, and, Third Department of Internal Medicine, Justus-Liebig-Universität, Giessen, Germany. triantafyllos.chavakis@innere.med.uni-giessen.de
authors.forename: T
authors.initials: T
authors.lastname: Chavakis
authors.forename: S M
authors.initials: SM
authors.lastname: Kanse
authors.forename: R A
authors.initials: RA
authors.lastname: Pixley
authors.forename: A E
authors.initials: AE
authors.lastname: May
authors.forename: I
authors.initials: I
authors.lastname: Isordia-Salas
authors.forename: R W
authors.initials: RW
authors.lastname: Colman
authors.forename: K T
authors.initials: KT
authors.lastname: Preissner
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001667:Binding, Competitive / Q000187:drug effects; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D005260:Female; D005340:Fibrinogen / Q000378:metabolism; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D020014:K562 Cells; D019679:Kininogen, High-Molecular-Weight / Q000737:chemistry / Q000494:pharmacology*; D007962:Leukocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D016169:Lymphocyte Function-Associated Antigen-1 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D016177:Macrophage-1 Antigen / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D020556:Neutrophil Infiltration / Q000187:drug effects; D010455:Peptides / Q000494:pharmacology*; D010538:Peritonitis / Q000139:chemically induced / Q000473:pathology / Q000517:prevention & control; D010957:Plasmids / Q000235:genetics; D013864:Thioglycolates / Q000008:administration & dosage; D020298:U937 Cells
chemical_list: D000911:Antibodies, Monoclonal; D019679:Kininogen, High-Molecular-Weight; D016169:Lymphocyte Function-Associated Antigen-1; D016177:Macrophage-1 Antigen; D010455:Peptides; D013864:Thioglycolates; D018799:Intercellular Adhesion Molecule-1; D005340:Fibrinogen