title: Establishment of neutralizing rat monoclonal antibodies for fibroblast growth factor-2.
abstract: Fibroblast growth factor-2 (FGF-2) plays a critical role in endothelial survival, proliferation, and angiogenesis and is localized on the cell membrane by binding to heparan sulfate proteoglycans. Here we established a neutralizing monoclonal antibody, 1B9B9, against FGF-2 using the rat medial iliac lymph node method. 1B9B9 blocked the binding of FGF-2 to its receptor, inhibiting FGF-2-induced proliferation and corresponding downstream signaling in endothelial cells. Treatment of human umbilical vein endothelial cells with 1B9B9 reduced the basal phosphorylation levels of Akt and MAPK. Furthermore, continued treatment with 1B9B9 induced cell death by apoptosis. Compared with FGF-2 knockdown, 1B9B9 significantly reduced cell survival. In addition, the combination of FGF-2 siRNA and 1B9B9 showed a synergistic effect. The data indicate that 1B9B9 established by the rat iliac lymph node method is a fully compatible neutralizing antibody.
authors.affiliation: 1 Department of Pharmacology, Osaka City University Medical School , Osaka, Japan .
authors.forename: Masako
authors.initials: M
authors.lastname: Tanaka
authors.forename: Maki
authors.initials: M
authors.lastname: Yamaguchi
authors.forename: Masayuki
authors.initials: M
authors.lastname: Shiota
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Kawamoto
authors.forename: Katsuyuki
authors.initials: K
authors.lastname: Takahashi
authors.forename: Azusa
authors.initials: A
authors.lastname: Inagaki
authors.forename: Mayuko
authors.initials: M
authors.lastname: Osada-Oka
authors.forename: Akihito
authors.initials: A
authors.lastname: Harada
authors.forename: Hideki
authors.initials: H
authors.lastname: Wanibuchi
authors.forename: Yasukatsu
authors.initials: Y
authors.lastname: Izumi
authors.forename: Katsuyuki
authors.initials: K
authors.lastname: Miura
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Iwao
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Ohkawa
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D057134:Antibodies, Neutralizing / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D042783:Endothelial Cells / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D016222:Fibroblast Growth Factor 2 / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D008856:Microscopy, Fluorescence; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D051058:Oncogene Protein v-akt / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D034741:RNA, Small Interfering; D016222:Fibroblast Growth Factor 2; D051058:Oncogene Protein v-akt; D020928:Mitogen-Activated Protein Kinases