title: Monoclonal antibodies to vascular endothelial growth factor-D block its interactions with both VEGF receptor-2 and VEGF receptor-3.
abstract: Vascular endothelial growth factor-D (VEGF-D), the most recently discovered mammalian member of the VEGF family, is an angiogenic protein that activates VEGF receptor-2 (VEGFR-2/Flk1/KDR) and VEGFR-3 (Flt4). These receptor tyrosine kinases, localized on vascular and lymphatic endothelial cells, signal for angiogenesis and lymphangiogenesis. VEGF-D consists of a central receptor-binding VEGF homology domain (VHD) and N-terminal and C-terminal propeptides that are cleaved from the VHD to generate a mature, bioactive form consisting of dimers of the VHD. Here we report characterization of mAbs raised to the VHD of human VEGF-D in order to generate VEGF-D antagonists. The mAbs bind the fully processed VHD with high affinity and also bind unprocessed VEGF-D. We demonstrate, using bioassays for the binding and cross-linking of VEGFR-2 and VEGFR-3 and biosensor analysis with immobilized receptors, that one of the mAbs, designated VD1, is able to compete potently with mature VEGF-D for binding to both VEGFR-2 and VEGFR-3 for binding to mature VEGF-D. This indicates that the binding epitopes on VEGF-D for these two receptors may be in close proximity. Furthermore, VD1 blocks the mitogenic response of human microvascular endothelial cells to VEGF-D. The anti-(VEGF-D) mAbs raised to the bioactive region of this growth factor will be powerful tools for analysis of the biological functions of VEGF-D.
authors.affiliation: Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Victoria, Australia. marc.achen@ludwig.edu.au
authors.forename: M G
authors.initials: MG
authors.lastname: Achen
authors.forename: S
authors.initials: S
authors.lastname: Roufail
authors.forename: T
authors.initials: T
authors.lastname: Domagala
authors.forename: B
authors.initials: B
authors.lastname: Catimel
authors.forename: E C
authors.initials: EC
authors.lastname: Nice
authors.forename: D M
authors.initials: DM
authors.lastname: Geleick
authors.forename: R
authors.initials: R
authors.lastname: Murphy
authors.forename: A M
authors.initials: AM
authors.lastname: Scott
authors.forename: C
authors.initials: C
authors.lastname: Caesar
authors.forename: T
authors.initials: T
authors.lastname: Makinen
authors.forename: K
authors.initials: K
authors.lastname: Alitalo
authors.forename: S A
authors.initials: SA
authors.lastname: Stacker
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D001483:Base Sequence; D002455:Cell Division / Q000276:immunology; D002478:Cells, Cultured; D017931:DNA Primers; D016228:Endothelial Growth Factors / Q000276:immunology / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology / Q000276:immunology; D018604:Epitope Mapping; D006801:Humans; D020794:Receptor Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors* / Q000378:metabolism; D017978:Receptors, Growth Factor / Q000037:antagonists & inhibitors* / Q000378:metabolism; D040262:Receptors, Vascular Endothelial Growth Factor; D042643:Vascular Endothelial Growth Factor D; D040321:Vascular Endothelial Growth Factor Receptor-3
chemical_list: D000911:Antibodies, Monoclonal; D017931:DNA Primers; D016228:Endothelial Growth Factors; D011956:Receptors, Cell Surface; D017978:Receptors, Growth Factor; D042643:Vascular Endothelial Growth Factor D; D020794:Receptor Protein-Tyrosine Kinases; D040262:Receptors, Vascular Endothelial Growth Factor; D040321:Vascular Endothelial Growth Factor Receptor-3