Role of neutrophil elastase in LTB4‐induced neutrophil transmigration in vivo assessed with a specific inhibitor and neutrophil elastase deficient mice

• Published in: British journal of pharmacology 2007-07, Vol.151 (5), p.628-637
• Main Authors: Young, R E, Voisin, M‐B, Wang, S, Dangerfield, J, Nourshargh, S
• Format: Article
• Language: English
• Subjects: Animals; basement membrane; Basement Membrane - drug effects; Basement Membrane - metabolism; Biological and medical sciences; Bone Marrow Cells - drug effects; Bone Marrow Cells - metabolism; Cell Movement - drug effects; cell trafficking; Chemotaxis, Leukocyte - drug effects; Enzyme Inhibitors - pharmacology; Glycine - analogs & derivatives; Glycine - pharmacology; inflammation; intravital microscopy; Leukocyte Elastase - antagonists & inhibitors; Leukocyte Elastase - deficiency; Leukocyte Elastase - physiology; Leukotriene B4 - pharmacology; Male; Medical sciences; Membrane Proteins - biosynthesis; Mice; Mice, Inbred C57BL; Mice, Knockout; Microscopy, Confocal; neutrophil elastase; Neutrophils - drug effects; Neutrophils - enzymology; Pharmacology. Drug treatments; Platelet Activating Factor - metabolism; Research Papers; Sulfonamides - pharmacology; Venules - drug effects; Venules - metabolism
• ISSN: 0007-1188; 1476-5381
• DOI: 10.1038/sj.bjp.0707267

Background and purpose: The serine protease neutrophil elastase (NE) appears to regulate inflammatory responses at multiple levels but its role in leukocyte transmigration in vivo remains unclear. The present study aimed to address this issue by using both an NE inhibitor (ONO‐5046) and NE deficient (NE−/−) mice. Experimental approach: A number of inflammatory mediators (LTB4, KC and PAF) were investigated in vitro for their ability to stimulate the release and the surface expression of NE by neutrophils. In addition, the role of NE in leukocyte migration elicited by topical LTB4 was investigated in vivo in mouse cremasteric venules as observed by intravital microscopy. Key results: Amongst the mediators tested in vitro, LTB4 was found to be a highly potent and efficacious inducer of NE cell surface expression on murine neutrophils. Furthermore, in wild‐type mice (WT), LTB4‐induced leukocyte transmigration was reduced by intravenous ONO‐5046 (66% inhibition), an effect that appeared to occur at the level of the perivascular basement membrane. Interestingly, LTB4‐induced responses were normal in NE−/− mice and, while ONO‐5046 had no inhibitory effect in these animals, the broad‐spectrum serine protease inhibitor aprotinin suppressed leukocyte transmigration in both WT and NE−/− mice. Conclusions and implications: The findings demonstrate the potent ability of LTB4 to induce cell‐surface expression of NE and provide evidence for the involvement of NE in LTB4‐induced neutrophil transmigration in vivo. The results also suggest the existence of compensatory mechanisms in NE−/− mice, highlighting the added value of investigating pharmacological blockers in parallel with genetic deletion. British Journal of Pharmacology (2007) 151, 628–637; doi:10.1038/sj.bjp.0707267