title: Involvement of the VEGF receptor 3 in tubular morphogenesis demonstrated with a human anti-human VEGFR-3 monoclonal antibody that antagonizes receptor activation by VEGF-C.
abstract: In this report we utilize a novel antagonist antibody to the human VEGFR-3 to elucidate the role of this receptor in in vitro tubular morphogenesis of bovine and human endothelial cells (EC cells) induced by VEGF-C. The antibody hF4-3C5 was obtained by panning a human phage display library on soluble human VEGFR-3. The binding affinity constant of hF4-3C5 significantly exceeds that of the interaction of VEGFR-3 with VEGF-C. hF4-3C5 strongly inhibits the binding of soluble VEGFR-3 to immobilized VEGF-C and abolishes the VEGF-C-mediated mitogenic response of cells that expresses a chimeric human VEGFR-3-cFMS receptor. In fluorescence experiments, hF4-3C5 reactivity is observed with human lymphatic endothelial cells (LECs) and human umbilical vein endothelial cells (HUVECs). Binding of hF4-3C5 shows that about half of bovine aortic endothelial (BAE) cells express VEGFR-3 and cells in this subpopulation are primarily responsible for the chemotactic response to the mature form of VEGF-C (VEGF-C(DeltaNDeltaC)). This response was strongly inhibited by the addition of hF4-3C5. In vitro tube formation by BAE cells induced by VEGF-C(DeltaNDeltaC) was reduced by greater than 60% by hF4-3C5 whereas the response to VEGF(165) was unaffected. Addition of hF4-3C5 together with an antagonist antibody to VEGFR-2 completely abolished the response to VEGF-C(DeltaNDeltaC). Similar results were obtained with HUVECs. Together, these findings point to a role for VEGFR-3 in vascular tubular morphogenesis and highlight the utility of hF4-3C5 as a tool for the investigation of the biology of VEGFR-3.
authors.affiliation: ImClone Systems Incorporated, New York, NY 10014, USA.
authors.forename: Kris
authors.initials: K
authors.lastname: Persaud
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Tille
authors.forename: Meilin
authors.initials: M
authors.lastname: Liu
authors.forename: Zhenping
authors.initials: Z
authors.lastname: Zhu
authors.forename: Xenia
authors.initials: X
authors.lastname: Jimenez
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Pereira
authors.forename: Hua-Quan
authors.initials: HQ
authors.lastname: Miao
authors.forename: Laura A
authors.initials: LA
authors.lastname: Brennan
authors.forename: Larry
authors.initials: L
authors.lastname: Witte
authors.forename: Michael S
authors.initials: MS
authors.lastname: Pepper
authors.forename: Bronislaw
authors.initials: B
authors.lastname: Pytowski
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D001011:Aorta / Q000166:cytology; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D002633:Chemotaxis / Q000187:drug effects; D002999:Clone Cells; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000254:growth & development*; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D008208:Lymphatic System / Q000166:cytology; D051379:Mice; D008856:Microscopy, Fluorescence; D009024:Morphogenesis; D041681:NIH 3T3 Cells; D019151:Peptide Library; D011994:Recombinant Proteins / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology; D042582:Vascular Endothelial Growth Factor C / Q000378:metabolism* / Q000494:pharmacology; D040321:Vascular Endothelial Growth Factor Receptor-3 / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D017077:Culture Media, Conditioned; D019151:Peptide Library; D011994:Recombinant Proteins; D042582:Vascular Endothelial Growth Factor C; D040321:Vascular Endothelial Growth Factor Receptor-3