Soluble factor(s) released from neutrophils activates endothelial cell matrix metalloproteinase-2

Jess D. Schwartz, Sara Monea, Stuart G. Marcus, Sundeep Patel, Kenneth Eng, Aubrey C. Galloway, Paolo Mignatti, Peter Shamamian

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Objective. Polymorphonuclear leukocyte (PMN) infiltration and microvascular injury are hallmarks of the tissue remodeling associated with multiple organ failure. These processes require the concerted action of various proteolytic enzymes, including serine and matrix metalloproteinases (MMPs). Matrix metalloproteinase-2 (MMP-2) plays an important role in the turnover of various ECM components, including type IV collagen, fibronectin, and gelatins. Like all MMPs, MMP-2 is secreted as an inactive zymogen (proMMP2) and activated extracellularly by limited proteolytic cleavage. The physiologic mechanism(s) of proMMP-2 activation remains unclear. This study was designed to characterize the effect of PMNs on the activation of proMMP- 2 produced by endothelial cells. Methods. PMNs and human umbilical vein endothelial cells (HUVECs) were grown either separately or together for 2-16 h. To evaluate the role of cell-cell contact, cocultures were also established in which the two cell types were separated by a semipermeable polycarbonate membrane. Alternatively, PMN-conditioned medium was added to HUVEC cultures with or without various proteinase inhibitors (aprotinin, 1.10-phenanthroline, Batimastat, E-64, eglin c peptide, or pepstatin A). After incubation, the culture supernatants were analyzed by gelatin zymography to characterize the gelatinases. Results. HUVECs produce MMP-2 in its inactive (72 kDa) form. PMNs produce high levels of MMP-9 (gelatinase B, 92 kDa) but no MMP-2. Coculture of PMNs with or addition of PMN-conditioned medium to HUVECs results in the production of active (62 kDa) MMP-2. ProMMP- 2 activation by PMN-conditioned medium is not blocked by inhibitors of plasmin, cysteine-, acid-, or metalloproteinases. Conclusion. PMNs release a soluble factor that activates endothelial cell MMP-2 through a novel mechanism independent of cell-cell contact and not attributable to the activities of plasmin, cysteine-, acid-, or metalloproteinases. These findings may provide insight into the tissue remodeling that accompanies PMN- mediated microvascular injury.

Original languageEnglish (US)
Pages (from-to)79-85
Number of pages7
JournalJournal of Surgical Research
Volume76
Issue number1
DOIs
StatePublished - Apr 1998
Externally publishedYes

Fingerprint

Matrix Metalloproteinase 2
Neutrophils
Endothelial Cells
Human Umbilical Vein Endothelial Cells
Conditioned Culture Medium
polycarbonate
Matrix Metalloproteinase 9
Metalloproteases
Gelatin
Coculture Techniques
Matrix Metalloproteinases
Cysteine
Peptide Hydrolases
Gelatinases
Antifibrinolytic Agents
Enzyme Precursors
Aprotinin
Acids
Phenanthrolines
Collagen Type IV

Keywords

  • Matrix metalloproteinases
  • MMP-2
  • Neutrophils
  • Proteolytic activation

ASJC Scopus subject areas

  • Surgery

Cite this

Soluble factor(s) released from neutrophils activates endothelial cell matrix metalloproteinase-2. / Schwartz, Jess D.; Monea, Sara; Marcus, Stuart G.; Patel, Sundeep; Eng, Kenneth; Galloway, Aubrey C.; Mignatti, Paolo; Shamamian, Peter.

In: Journal of Surgical Research, Vol. 76, No. 1, 04.1998, p. 79-85.

Research output: Contribution to journalArticle

Schwartz, JD, Monea, S, Marcus, SG, Patel, S, Eng, K, Galloway, AC, Mignatti, P & Shamamian, P 1998, 'Soluble factor(s) released from neutrophils activates endothelial cell matrix metalloproteinase-2', Journal of Surgical Research, vol. 76, no. 1, pp. 79-85. https://doi.org/10.1006/jsre.1998.5294
Schwartz, Jess D. ; Monea, Sara ; Marcus, Stuart G. ; Patel, Sundeep ; Eng, Kenneth ; Galloway, Aubrey C. ; Mignatti, Paolo ; Shamamian, Peter. / Soluble factor(s) released from neutrophils activates endothelial cell matrix metalloproteinase-2. In: Journal of Surgical Research. 1998 ; Vol. 76, No. 1. pp. 79-85.
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abstract = "Objective. Polymorphonuclear leukocyte (PMN) infiltration and microvascular injury are hallmarks of the tissue remodeling associated with multiple organ failure. These processes require the concerted action of various proteolytic enzymes, including serine and matrix metalloproteinases (MMPs). Matrix metalloproteinase-2 (MMP-2) plays an important role in the turnover of various ECM components, including type IV collagen, fibronectin, and gelatins. Like all MMPs, MMP-2 is secreted as an inactive zymogen (proMMP2) and activated extracellularly by limited proteolytic cleavage. The physiologic mechanism(s) of proMMP-2 activation remains unclear. This study was designed to characterize the effect of PMNs on the activation of proMMP- 2 produced by endothelial cells. Methods. PMNs and human umbilical vein endothelial cells (HUVECs) were grown either separately or together for 2-16 h. To evaluate the role of cell-cell contact, cocultures were also established in which the two cell types were separated by a semipermeable polycarbonate membrane. Alternatively, PMN-conditioned medium was added to HUVEC cultures with or without various proteinase inhibitors (aprotinin, 1.10-phenanthroline, Batimastat, E-64, eglin c peptide, or pepstatin A). After incubation, the culture supernatants were analyzed by gelatin zymography to characterize the gelatinases. Results. HUVECs produce MMP-2 in its inactive (72 kDa) form. PMNs produce high levels of MMP-9 (gelatinase B, 92 kDa) but no MMP-2. Coculture of PMNs with or addition of PMN-conditioned medium to HUVECs results in the production of active (62 kDa) MMP-2. ProMMP- 2 activation by PMN-conditioned medium is not blocked by inhibitors of plasmin, cysteine-, acid-, or metalloproteinases. Conclusion. PMNs release a soluble factor that activates endothelial cell MMP-2 through a novel mechanism independent of cell-cell contact and not attributable to the activities of plasmin, cysteine-, acid-, or metalloproteinases. These findings may provide insight into the tissue remodeling that accompanies PMN- mediated microvascular injury.",
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AU - Monea, Sara

AU - Marcus, Stuart G.

AU - Patel, Sundeep

AU - Eng, Kenneth

AU - Galloway, Aubrey C.

AU - Mignatti, Paolo

AU - Shamamian, Peter

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N2 - Objective. Polymorphonuclear leukocyte (PMN) infiltration and microvascular injury are hallmarks of the tissue remodeling associated with multiple organ failure. These processes require the concerted action of various proteolytic enzymes, including serine and matrix metalloproteinases (MMPs). Matrix metalloproteinase-2 (MMP-2) plays an important role in the turnover of various ECM components, including type IV collagen, fibronectin, and gelatins. Like all MMPs, MMP-2 is secreted as an inactive zymogen (proMMP2) and activated extracellularly by limited proteolytic cleavage. The physiologic mechanism(s) of proMMP-2 activation remains unclear. This study was designed to characterize the effect of PMNs on the activation of proMMP- 2 produced by endothelial cells. Methods. PMNs and human umbilical vein endothelial cells (HUVECs) were grown either separately or together for 2-16 h. To evaluate the role of cell-cell contact, cocultures were also established in which the two cell types were separated by a semipermeable polycarbonate membrane. Alternatively, PMN-conditioned medium was added to HUVEC cultures with or without various proteinase inhibitors (aprotinin, 1.10-phenanthroline, Batimastat, E-64, eglin c peptide, or pepstatin A). After incubation, the culture supernatants were analyzed by gelatin zymography to characterize the gelatinases. Results. HUVECs produce MMP-2 in its inactive (72 kDa) form. PMNs produce high levels of MMP-9 (gelatinase B, 92 kDa) but no MMP-2. Coculture of PMNs with or addition of PMN-conditioned medium to HUVECs results in the production of active (62 kDa) MMP-2. ProMMP- 2 activation by PMN-conditioned medium is not blocked by inhibitors of plasmin, cysteine-, acid-, or metalloproteinases. Conclusion. PMNs release a soluble factor that activates endothelial cell MMP-2 through a novel mechanism independent of cell-cell contact and not attributable to the activities of plasmin, cysteine-, acid-, or metalloproteinases. These findings may provide insight into the tissue remodeling that accompanies PMN- mediated microvascular injury.

AB - Objective. Polymorphonuclear leukocyte (PMN) infiltration and microvascular injury are hallmarks of the tissue remodeling associated with multiple organ failure. These processes require the concerted action of various proteolytic enzymes, including serine and matrix metalloproteinases (MMPs). Matrix metalloproteinase-2 (MMP-2) plays an important role in the turnover of various ECM components, including type IV collagen, fibronectin, and gelatins. Like all MMPs, MMP-2 is secreted as an inactive zymogen (proMMP2) and activated extracellularly by limited proteolytic cleavage. The physiologic mechanism(s) of proMMP-2 activation remains unclear. This study was designed to characterize the effect of PMNs on the activation of proMMP- 2 produced by endothelial cells. Methods. PMNs and human umbilical vein endothelial cells (HUVECs) were grown either separately or together for 2-16 h. To evaluate the role of cell-cell contact, cocultures were also established in which the two cell types were separated by a semipermeable polycarbonate membrane. Alternatively, PMN-conditioned medium was added to HUVEC cultures with or without various proteinase inhibitors (aprotinin, 1.10-phenanthroline, Batimastat, E-64, eglin c peptide, or pepstatin A). After incubation, the culture supernatants were analyzed by gelatin zymography to characterize the gelatinases. Results. HUVECs produce MMP-2 in its inactive (72 kDa) form. PMNs produce high levels of MMP-9 (gelatinase B, 92 kDa) but no MMP-2. Coculture of PMNs with or addition of PMN-conditioned medium to HUVECs results in the production of active (62 kDa) MMP-2. ProMMP- 2 activation by PMN-conditioned medium is not blocked by inhibitors of plasmin, cysteine-, acid-, or metalloproteinases. Conclusion. PMNs release a soluble factor that activates endothelial cell MMP-2 through a novel mechanism independent of cell-cell contact and not attributable to the activities of plasmin, cysteine-, acid-, or metalloproteinases. These findings may provide insight into the tissue remodeling that accompanies PMN- mediated microvascular injury.

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