Inhibition of angiogenesis by antibody blocking the action of proangiogenic high-molecular-weight kininogen

J Thromb Haemost. 2003 Jan;1(1):164-70. doi: 10.1046/j.1538-7836.2003.00025.x.


Previously we demonstrated that domain 5 (D5) of high-molecular-weight kininogen (HK) inhibits neovascularization in the chicken chorioallantoic membrane (CAM) assay and further found that kallikrein cleaved HK (HKa) inhibited FGF2-and VEGF-induced neovascularization, and thus was antiangiogenic. In this study, we sought to demonstrate whether uncleaved HK stimulates neovascularization and thus is proangiogenic. The chick chorioallantoic membrane was used as an in ovo assay of angiogenesis. Low-molecular-weight kininogen stimulates angiogenesis, indicating that D5 is not involved. Bradykinin stimulates neovascularization equally to HK and LK and is likely to be responsible for the effect of HK. A murine monoclonal antibody to HK (C11C1) also recognizes a similar component in chicken plasma as detected by surface plasmon resonance. Angiogenesis induced by FGF2 and VEGF is inhibited by this monoclonal antibody and is a more potent inhibitor of neovascularization induced by VEGF than an integrin alphavbeta3 antibody (LM 609). Our postulate that C11C1 inhibits the stimulation of angiogenesis by HK was confirmed when either C11C1 or D5 completely inhibited angiogenesis in the CAM induced by HK. Growth of human fibrosarcoma (HT-1080) on the CAM was inhibited by GST-D5 and C11C1. These results indicate HK is proangiogenic probably by releasing bradykinin and that a monoclonal antibody directed to HK could serve as an antiangiogenic agent with a potential for inhibiting tumor angiogenesis and other angiogenesis-mediated disorders.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Allantois / blood supply
  • Angiogenesis Inhibitors / pharmacology*
  • Animals
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / pharmacology*
  • Bradykinin / pharmacology
  • Chick Embryo
  • Chorion / blood supply
  • Dose-Response Relationship, Drug
  • Endothelial Growth Factors / pharmacology
  • Fibroblast Growth Factor 2 / pharmacology
  • Fibrosarcoma / metabolism
  • Glutathione Transferase / genetics
  • Glutathione Transferase / pharmacology
  • Humans
  • Intercellular Signaling Peptides and Proteins / pharmacology
  • Kininogen, High-Molecular-Weight / antagonists & inhibitors*
  • Kininogen, High-Molecular-Weight / immunology
  • Kininogen, High-Molecular-Weight / pharmacology
  • Kininogen, Low-Molecular-Weight / pharmacology
  • Lymphokines / pharmacology
  • Neovascularization, Physiologic / immunology*
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / pharmacology
  • Surface Plasmon Resonance
  • Tumor Cells, Cultured
  • Vascular Endothelial Growth Factor A
  • Vascular Endothelial Growth Factors


  • Angiogenesis Inhibitors
  • Antibodies, Monoclonal
  • Endothelial Growth Factors
  • Intercellular Signaling Peptides and Proteins
  • Kininogen, High-Molecular-Weight
  • Kininogen, Low-Molecular-Weight
  • Lymphokines
  • Recombinant Fusion Proteins
  • Vascular Endothelial Growth Factor A
  • Vascular Endothelial Growth Factors
  • Fibroblast Growth Factor 2
  • Glutathione Transferase
  • Bradykinin