TY - JOUR
T1 - Establishment of neutralizing rat monoclonal antibodies for fibroblast growth factor-2
AU - Tanaka, Masako
AU - Yamaguchi, Maki
AU - Shiota, Masayuki
AU - Kawamoto, Yukiko
AU - Takahashi, Katsuyuki
AU - Inagaki, Azusa
AU - Osada-Oka, Mayuko
AU - Harada, Akihito
AU - Wanibuchi, Hideki
AU - Izumi, Yasukatsu
AU - Miura, Katsuyuki
AU - Iwao, Hiroshi
AU - Ohkawa, Yasuyuki
PY - 2014/8/1
Y1 - 2014/8/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84907033952&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84907033952&partnerID=8YFLogxK
U2 - 10.1089/mab.2013.0085
DO - 10.1089/mab.2013.0085
M3 - Article
C2 - 25171006
AN - SCOPUS:84907033952
SN - 2167-9436
VL - 33
SP - 261
EP - 269
JO - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
JF - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
IS - 4
ER -