Proinflammatory mediators stimulate neutrophil-directed angiogenesis.
McCourt, M Wang, J H Sookhai, S Redmond, H P
McCourt, M
Wang, J H
Sookhai, S
Redmond, H P
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Date
2012-02-03T15:13:16Z
Date Submitted
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Subject Mesh
Antigens, CD95/physiology
Cells, Cultured
Endothelial Growth Factors/physiology
Endothelium, Vascular/*immunology
Humans
Inflammation Mediators/*physiology
Interleukin-6/physiology
Lipopolysaccharides/immunology
Lymphokines/physiology
Neovascularization, Pathologic/*immunology
Neutrophils/*immunology
Tumor Necrosis Factor-alpha/physiology
Vascular Endothelial Growth Factor A
Vascular Endothelial Growth Factors
Cells, Cultured
Endothelial Growth Factors/physiology
Endothelium, Vascular/*immunology
Humans
Inflammation Mediators/*physiology
Interleukin-6/physiology
Lipopolysaccharides/immunology
Lymphokines/physiology
Neovascularization, Pathologic/*immunology
Neutrophils/*immunology
Tumor Necrosis Factor-alpha/physiology
Vascular Endothelial Growth Factor A
Vascular Endothelial Growth Factors
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Abstract
BACKGROUND: Vascular endothelial growth factor (VEGF; vascular permeability factor) is one of the most potent proangiogenic cytokines, and it plays a central role in mediating the process of angiogenesis or new blood vessel formation. Neutrophils (PMNs) recently have been shown to produce VEGF. HYPOTHESIS: The acute inflammatory response is a potent stimulus for PMN-directed angiogenesis. METHODS: Neutrophils were isolated from healthy volunteers and stimulated with lipopolysaccharide (LPS), tumor necrosis factor alpha (TNF-alpha), interleukin 6 (IL-6), and anti-human Fas monoclonal antibody. Culture supernatants were assayed for VEGF using enzyme-linked immunosorbent assays. Culture supernatants from LPS- and TNF-alpha-stimulated PMNs were then added to human umbilical vein endothelial cells and human microvessel endothelial cells and assessed for endothelial cell proliferation using 5-bromodeoxyuridine labeling. Tubule formation was also assessed on MATRIGEL basement membrane matrix. Neutrophils were lysed to measure total VEGF release, and VEGF expression was detected using Western blot analysis. RESULTS: Lipopolysaccharide and TNF-alpha stimulation resulted in significantly increased release of PMN VEGF (532+/-49 and 484+/-80 pg/mL, respectively; for all, presented as mean +/- SEM) compared with control experiments (32+/-4 pg/mL). Interleukin 6 and Fas had no effect. Culture supernatants from LPS- and TNF-alpha-stimulated PMNs also resulted in significant increases (P<.005) in macrovascular and microvascular endothelial cell proliferation and tubule formation. Adding anti-human VEGF-neutralizing polyclonal antibody to stimulated PMN supernatant inhibited these effects. Total VEGF release following cell lysis and Western blot analysis suggests that the VEGF is released from an intracellular store. CONCLUSION: Activated human PMNs are directly angiogenic by releasing VEGF, and this has important implications for inflammation, capillary leak syndrome, wound healing, and tumor growth.
Language
eng
ISSN
0004-0010 (Print)
0004-0010 (Linking)
0004-0010 (Linking)
eISSN
ISBN
DOI
PMID
10593330