title: Elastase release by transmigrating neutrophils deactivates endothelial-bound SDF-1alpha and attenuates subsequent T lymphocyte transendothelial migration.
abstract: Leukocyte trafficking to sites of inflammation follows a defined temporal pattern, and evidence suggests that initial neutrophil transendothelial migration modifies endothelial cell phenotype. We tested the hypothesis that preconditioning of human umbilical vein endothelial cells (HUVEC) by neutrophils would also modify the subsequent transendothelial migration of T lymphocytes across cytokine-stimulated HUVEC in an in vitro flow assay. Using fluorescence microscopy, preconditioning of HUVEC by neutrophils was observed to significantly reduce the extent of subsequent stromal cell-derived factor-1alpha (SDF-1alpha [CXCL12])-mediated T lymphocyte transendothelial migration, without reducing accumulation. In contrast, recruitment of a second wave of neutrophils was unaltered. Conditioned medium harvested after transendothelial migration of neutrophils or supernatants from stimulated neutrophils mediated a similar blocking effect, which was negated using a specific neutrophil elastase inhibitor. Furthermore, T lymphocyte transendothelial migration was inhibited by treatment of HUVEC with purified neutrophil elastase, which selectively cleaved the amino terminus of HUVEC-bound SDF-1alpha, which is required for its chemotactic activity. The reduction in T lymphocyte transendothelial migration was not observed using a different chemokine, ELC (CCL19), and was not reversed by replenishment of SDF-1alpha, indicating endothelial retention of the inactivated chemokine. In summary, transmigrating neutrophils secrete localized elastase that is protected from plasma inhibitors, and thereby modulate trafficking of other leukocyte subsets by altering the endothelial-associated chemotactic activities.
authors.affiliation: Department of Pathology, Center for Excellence in Vascular Biology, Brigham and Women's Hospital, Harvard Medical School, 77 Ave. Louis Pasteur NRB 752P, Boston, MA 02115, USA.
authors.forename: Ravi M
authors.initials: RM
authors.lastname: Rao
authors.forename: Travis V
authors.initials: TV
authors.lastname: Betz
authors.forename: Deanna J
authors.initials: DJ
authors.lastname: Lamont
authors.forename: Michael B
authors.initials: MB
authors.lastname: Kim
authors.forename: Sunil K
authors.initials: SK
authors.lastname: Shaw
authors.forename: Richard M
authors.initials: RM
authors.lastname: Froio
authors.forename: Françoise
authors.initials: F
authors.lastname: Baleux
authors.forename: Fernando
authors.initials: F
authors.lastname: Arenzana-Seisdedos
authors.forename: Ronen
authors.initials: R
authors.lastname: Alon
authors.forename: Francis W
authors.initials: FW
authors.lastname: Luscinskas
mesh_terms: D002450:Cell Communication; D002465:Cell Movement; D002478:Cells, Cultured; D054377:Chemokine CXCL12; D019743:Chemokines, CXC / Q000502:physiology*; D015936:Complement C5a / Q000494:pharmacology; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology; D016209:Interleukin-8 / Q000494:pharmacology; D019272:Leukocyte Elastase / Q000502:physiology*; D009504:Neutrophils / Q000502:physiology*; D013601:T-Lymphocytes / Q000502:physiology*
chemical_list: C515255:CXCL12 protein, human; D054377:Chemokine CXCL12; D019743:Chemokines, CXC; D007375:Interleukin-1; D016209:Interleukin-8; D015936:Complement C5a; D019272:Leukocyte Elastase