title: Role of gelatinase B and elastase in human polymorphonuclear neutrophil migration across basement membrane.
abstract: Polymorphonuclear neutrophil (PMN) migration across basement membrane is thought to be dependent on the degradation of membrane constituents. PMN gelatinase B, a metalloproteinase able to degrade type IV collagen, may be involved in this phenomenon. PMN gelatinase B is released in the extracellular medium as a latent proform and then activated, mainly by PMN elastase. We investigated the role of gelatinase B in PMN migration across a Matrigel basement membrane matrix coated onto a filter, in a Boyden chamber. The effects of gelatinase and elastase inhibitors on PMN migration in this system were tested. Chemokinesis of PMN was tested in the same Boyden chamber across a filter free of basement membrane. The agarose method was used to test the same inhibitors for effects on PMN chemotaxis. In both systems, FMLP 10(-7)M was used as a chemoattractant. Addition of 10(-8)M TIMP-1 (the preferential gelatinase B inhibitor) inhibited trans-basement membrane PMN migration by 52 +/- 6% (P<0.05), without affecting PMN chemokinesis, chemotaxis, or degranulation. Also, (Ala)(2) Pro Val chloromethyl ketone (AAPVCK) 100 micron, a specific elastase inhibitor, inhibited trans-basement membrane PMN migration by 51 +/- 8% (P<0.05), without affecting PMN chemokinesis, chemotaxis, or degranulation. The AAPVCK-TIMP combination led to a decrease in migration across Matrigel basement membrane (46 +/- 2%, P,0.05)similar to that seen with TIMP alone. AAPVCK was responsible for inhibition of gelatinase B activation, leading to a decrease in activated gelatinase from 14% to 2% of total gelatinase release (P<0.05). All these results strongly suggest that gelatinase B is a major factor of PMN migration across basement membrane and that elastase may contribute to this process by activating pro-gelatinase B.
authors.affiliation: Département de Physiologie, Service de Réanimation Médicale, Créteil, France.
authors.forename: C
authors.initials: C
authors.lastname: Delclaux
authors.forename: C
authors.initials: C
authors.lastname: Delacourt
authors.forename: M P
authors.initials: MP
authors.lastname: D'Ortho
authors.forename: V
authors.initials: V
authors.lastname: Boyer
authors.forename: C
authors.initials: C
authors.lastname: Lafuma
authors.forename: A
authors.initials: A
authors.lastname: Harf
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D001485:Basement Membrane / Q000276:immunology / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects / Q000276:immunology*; D002634:Chemotaxis, Leukocyte / Q000187:drug effects / Q000276:immunology; D003094:Collagen; D017364:Collagenases / Q000502:physiology*; D004338:Drug Combinations; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006023:Glycoproteins / Q000494:pharmacology; D006801:Humans; D006909:Hydroxyproline / Q000378:metabolism; D007797:Laminin; D019272:Leukocyte Elastase; D020780:Matrix Metalloproteinase 9; D061965:Matrix Metalloproteinase Inhibitors; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D018375:Neutrophil Activation; D009504:Neutrophils / Q000201:enzymology* / Q000502:physiology*; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors / Q000502:physiology*; D010618:Phenanthrolines / Q000494:pharmacology; D011509:Proteoglycans; D013450:Sulfones / Q000494:pharmacology; D019714:Tissue Inhibitor of Metalloproteinases
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D004338:Drug Combinations; D004791:Enzyme Inhibitors; D006023:Glycoproteins; D007797:Laminin; D061965:Matrix Metalloproteinase Inhibitors; D010618:Phenanthrolines; D011509:Proteoglycans; D013450:Sulfones; D019714:Tissue Inhibitor of Metalloproteinases; C059485:matrigel; C002010:4-(2-aminoethyl)benzenesulfonylfluoride; D009240:N-Formylmethionine Leucyl-Phenylalanine; D003094:Collagen; D010196:Pancreatic Elastase; D019272:Leukocyte Elastase; D017364:Collagenases; D020780:Matrix Metalloproteinase 9; D006909:Hydroxyproline