Migration of human granulocytes through reconstituted basement membrane is not dependent on matrix metalloproteinase-9 (MMP-9).
Transmigration of human granulocytes across a basal lamina equivalent was studied in vitro. Transwell inserts were coated with Matrigel, a reconstituted basement membrane. Granulocytes (2x10(6)) were applied to the upper chamber. As chemoattractant interleukin-8 (IL-8; 25 ng/ml) was added to the lower chamber. After 1 h of migration, cells were counted in the lower chamber. Specific hydroxamate inhibitors of MMPs (BB-3103, Ro 31-9790) or of serine proteases (Pefabloc, leupeptin) were added at various concentrations to both chambers before the start of migration. Additional experiments were performed with alpha(2)-macroglobulin, a natural inhibitor of MMPs and a monoclonal antibody which specifically blocks the activity of MMP-9. Migration of granulocytes through Matrigel could not be reduced significantly by any of the MMP inhibitors. A dose-dependent impairment of transmigration was only found with Pefabloc, however, this substance also induced severe morphological changes of the cells. The other inhibitor of serine proteases, leupeptin, did not influence migration at all.
Felkel
C
C
Department of Neurology, Georg August University, Robert-Koch-Str. 40, D-37075, Göttingen, Germany.
Schöll
U
U
Mäder
M
M
Schwartz
P
P
Felgenhauer
K
K
Hardeland
R
R
Beuche
W
W
Weber
F
F
D001485:Basement Membrane / Q000502:physiology*; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D002630:Chemotactic Factors / Q000494:pharmacology; D003094:Collagen / Q000494:pharmacology; D004338:Drug Combinations; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006098:Granulocytes / Q000502:physiology*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D016209:Interleukin-8 / Q000494:pharmacology; D007797:Laminin / Q000494:pharmacology; D020780:Matrix Metalloproteinase 9 / Q000502:physiology*; D061965:Matrix Metalloproteinase Inhibitors; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D011509:Proteoglycans / Q000494:pharmacology; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D000511:alpha-Macroglobulins / Q000494:pharmacology
D002630:Chemotactic Factors; D004338:Drug Combinations; D004791:Enzyme Inhibitors; D006877:Hydroxamic Acids; D016209:Interleukin-8; D007797:Laminin; D061965:Matrix Metalloproteinase Inhibitors; D011509:Proteoglycans; D015842:Serine Proteinase Inhibitors; D000511:alpha-Macroglobulins; C059485:matrigel; D009240:N-Formylmethionine Leucyl-Phenylalanine; D003094:Collagen; D020780:Matrix Metalloproteinase 9