Matsuzaki K, Yoshitake Y, Matuo Y, Sasaki H, Nishikawa K
Department of Biochemistry, Kanazawa Medical University, Ishikawa, Japan.
Proc Natl Acad Sci U S A. 1989 Dec;86(24):9911-5. doi: 10.1073/pnas.86.24.9911.
Two monoclonal antibodies (mAbs) against bovine heparin-binding growth factor II (HBGF-II)/basic fibroblast growth factor (bFGF) were obtained from mouse hybridoma cell lines. They were highly specific for bFGF from bovine, human, and mouse sources and did not cross-react with bovine heparin-binding growth factor I (HBGF-I)/acidic fibroblast growth factor (aFGF). The immunoglobulin class and subclass of these mAbs were IgG1, K. The apparent dissociation constant (Kd) for bFGF of these mAbs ranged from 10(-9) to 10(-10) M. One mAb (bFM-2) also cross-reacted with heat-inactivated bFGF, while the other mAb (bFM-1) did not, suggesting that bFM-1 recognized the conformation of the bFGF molecule necessary for its biological activity. These mAbs inhibited growth of cultured bovine capillary endothelial cells in both the presence and absence of exogenous bFGF, indicating the autocrine action of this growth factor in in vitro growth of these cells. On the other hand, injection of these hybridoma cell lines s.c. into the backs of athymic mice resulted in development of highly vascularized solid tumors and a sustained high level of anti-bFGF activity in the blood of the tumor-bearing mice. These findings suggest that bFGF is not essential as an autocrine or paracrine growth factor for angiogenesis in vivo. These mAbs should be useful in further studies on the physiological role and the conformation-function relationship of bFGF because they block its biological activity.
从鼠杂交瘤细胞系中获得了两种抗牛肝素结合生长因子II（HBGF-II）/碱性成纤维细胞生长因子（bFGF）的单克隆抗体（mAb）。它们对来自牛、人和小鼠来源的bFGF具有高度特异性，且不与牛肝素结合生长因子I（HBGF-I）/酸性成纤维细胞生长因子（aFGF）发生交叉反应。这些单克隆抗体的免疫球蛋白类别和亚类为IgG1、κ。这些单克隆抗体与bFGF的表观解离常数（Kd）范围为10^(-9)至10^(-10)M。一种单克隆抗体（bFM-2）也与热灭活的bFGF发生交叉反应，而另一种单克隆抗体（bFM-1）则不发生交叉反应，这表明bFM-1识别bFGF分子发挥其生物学活性所必需的构象。这些单克隆抗体在有和没有外源性bFGF的情况下均抑制培养的牛毛细血管内皮细胞的生长，表明该生长因子在这些细胞的体外生长中具有自分泌作用。另一方面，将这些杂交瘤细胞系皮下注射到无胸腺小鼠的背部，导致高度血管化实体瘤的形成以及荷瘤小鼠血液中持续高水平的抗bFGF活性。这些发现表明，bFGF作为体内血管生成的自分泌或旁分泌生长因子并非必不可少。这些单克隆抗体应有助于进一步研究bFGF的生理作用及其构象-功能关系，因为它们可阻断其生物学活性。
