摘要
本实验用Phast system电泳法,从大鼠视网膜神经节细胞诱向因子(retinal ganglion neuronotrophic factor,RGNTF)单克隆抗体(RGNTF-McAb)中,分离出一种免疫球蛋白lgG,免疫Balb/c小鼠,制备了抗RGNTF-McAb的独特型单克隆抗体。用MTT微量比色法,对该独特型单克隆抗体进行筛选。结果表明,独特型单克隆抗体G_4能特异性地和RGNTF单克隆抗体D_3结合,中和D_3对成视网膜细胞瘤(Rb)细胞的抑制作用;具有RGNTF的生物学效应,能促进Rb细胞的生长。用免疫组织化学方法证明,Rb细胞、大鼠胚胎视网膜节细胞、新生大鼠的视网膜节细胞都可对G_4独特型抗体发生阳性免疫反应,阳性颗粒较粗,主要分布在Rb细胞和视网膜节细胞的胞膜上,这些阳性颗粒可能是RGNTF的膜受体。所以我们认为,G_4单克隆抗体不仅是独特型抗体,而且是对位型的独特型抗体。它的产生对深入研究RGNTF的作用机制具有重要意义,亦有广阔的应用前景。
By immunizing Balb/c mice with immunoglobulin IgG which was isolated from the D_3 monoclonal antibody specific to the retinal ganglion neuronotrophic factor (RGNTF-McAb) with a Phast System gel electrophoretic method, we have developed anti-idiotypic antibodies against the RGNTF-McAb. A MTT colorimetric microassay designed to measure the effect of conditioned media from hybridomas containing antibodies on the growth activities of retinoblastoma (Rb) cells, was used to screen for idiotypic antibodies from these hybridomas. The results showed that G_4 hybridoma produced an anti-idiotypic antibody which could bind to the D_3 RGNTF-McAb and neutralize the inhibitory function of D3 on Rb cells, and possessed the trophic activity similar to RGNTF in promoting the growth activity of Rb cells. Immunohistochemical studies with anti-idiotypic antibody revealed that Rb cells and embryonic and neonatal rat retinal ganglion cells possessed coarse immunoreactive positive granules distributed on their cell surface membrane. These granules are identified as the receptors for RGNTF. Therefore, we concluded that the G_4 monoclonal antibody is not only an anti-idiotypic antibody but also an anti-paratopic antibody, It may be very useful in studying the mechanism of action of RGNTF and has a widespread applications in future.
出处
《解剖学报》
CAS
CSCD
北大核心
1992年第1期83-87,共5页
Acta Anatomica Sinica
关键词
独特型抗体
受体
视网膜神经
Anti-idiotypic antibody
Receptor
Retinal ganglion neuronotrophic factor
Monoclonal antibody
Retinoblastoma
