Generation of biologically active angiostatin kringle 1-3 by activated human neutrophils.
The contribution of polymorphonuclear neutrophils (PMN) to host defense and natural immunity extends well beyond their traditional role as professional phagocytes. In this study, we demonstrate that upon stimulation with proinflammatory stimuli, human PMN release enzymatic activities that, in vitro, generate bioactive angiostatin fragments from purified plasminogen. We also provide evidence that these angiostatin-like fragments, comprising kringle domain 1 to kringle domain 3 (kringle 1-3) of plasminogen, are generated as a byproduct of the selective proteolytic activity of neutrophil-secreted elastase. Remarkably, affinity-purified angiostatin kringle 1-3 fragments generated by neutrophils inhibited basic fibroblast growth factor plus vascular endothelial growth factor-induced endothelial cell proliferation in vitro, and both vascular endothelial growth factor-induced angiogenesis in the matrigel plug assay and fibroblast growth factor-induced angiogenesis in the chick embryo chorioallantoic membrane assay, in vivo. These results represent the first demonstration that biologically active angiostatin-like fragments can be generated by inflammatory human neutrophils. Because angiostatin is a potent inhibitor of angiogenesis, tumor growth, and metastasis, the data suggest that activated PMN not only act as potent effectors of inflammation, but might also play a critical role in the inhibition of angiogenesis in inflammatory diseases and tumors, by generation of a potent anti-angiogenic molecule.
Scapini
Patrizia
P
Section of General Pathology, Department of Pathology, University of Verona, Verona, Italy.
Nesi
Lorella
L
Morini
Monica
M
Tanghetti
Elena
E
Belleri
Mirella
M
Noonan
Douglas
D
Presta
Marco
M
Albini
Adriana
A
Cassatella
Marco A
MA
D043168:Angiostatins; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D002642:Chick Embryo; D017077:Culture Media, Conditioned; D016228:Endothelial Growth Factors / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D018082:Kringles / Q000502:physiology*; D019272:Leukocyte Elastase / Q000502:physiology; D008222:Lymphokines / Q000494:pharmacology; D018919:Neovascularization, Physiologic / Q000187:drug effects; D009504:Neutrophils / Q000378:metabolism*; D010446:Peptide Fragments / Q000096:biosynthesis* / Q000494:pharmacology; D010958:Plasminogen / Q000096:biosynthesis* / Q000378:metabolism / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D017077:Culture Media, Conditioned; D016228:Endothelial Growth Factors; D008222:Lymphokines; D010446:Peptide Fragments; D014409:Tumor Necrosis Factor-alpha; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D016222:Fibroblast Growth Factor 2; D007371:Interferon-gamma; D043168:Angiostatins; D010958:Plasminogen; D019272:Leukocyte Elastase