title: Production and characterization of a Tie2 agonist monoclonal antibody.
abstract: The Tie2 receptor and its known ligands, the angiopoietins, play a critical role in endothelial cell differentiation during the process of angiogenesis. Recent experimental observations indicate that the agonistic ligand, angiopoietin-1, can stimulate endothelial cell sprouting and act as a chemo-attractant in vitro and induce increased and enhanced angiogenesis both alone and in conjunction with vascular endothelial growth factor (VEGF) in vivo. Here, we present a monoclonal antibody (MAb), which binds to the extracellular portion of the Tie2 receptor and elicits similar agonist effects. Upon MAb binding to the native Tie2 receptor of cultured human umblical vein endothelial cells (HUVEC), there is a rapid increase in receptor autophosphorylation with a concomitant enhancement in the recruitment and association of the signalling intermediates Grb2 and SH-PTP2. The antibody further demonstrates functional activity in vascular tissues. In vitro, the antibody promotes the survival of cultured HUVEC and elicits a dose dependent outgrowth and branching of microvessels from cultured explants of rat aorta. When administered in vivo, the antibody enhances the vascularization of subcutaneous Matrigel implants in mice. Together these data suggest that the antibody is capable of acting as a surrogate ligand for Tie2 and further confirms the role of Tie2 in the differentiation of endothelial cells during angiogenesis.
authors.affiliation: SmithKline Beecham R & D, Departments of Oncology Research King of Prussia, Pennsylvania, USA. Michael_J_Hansbury@sbphrd.com
authors.forename: M J
authors.initials: MJ
authors.lastname: Hansbury
authors.forename: R F
authors.initials: RF
authors.lastname: Nicosia
authors.forename: W H
authors.initials: WH
authors.lastname: Zhu
authors.forename: S J
authors.initials: SJ
authors.lastname: Holmes
authors.forename: J D
authors.initials: JD
authors.lastname: Winkler
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000378:metabolism / Q000494:pharmacology; D001011:Aorta / Q000254:growth & development; D002454:Cell Differentiation; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D005260:Female; D006801:Humans; D008024:Ligands; D051379:Mice; D008810:Mice, Inbred C57BL; D009363:Neoplasm Proteins / Q000819:agonists* / Q000276:immunology* / Q000502:physiology; D018919:Neovascularization, Physiologic; D009924:Organ Culture Techniques; D010766:Phosphorylation; D011518:Proto-Oncogene Proteins; D051381:Rats; D042787:Receptor, TIE-2
chemical_list: D000911:Antibodies, Monoclonal; D008024:Ligands; C105373:MEN1 protein, human; D009363:Neoplasm Proteins; D011518:Proto-Oncogene Proteins; D042787:Receptor, TIE-2