Lan Hai-Nan, Jiang Hai-Long, Li Wei, Wu Tian-Cheng, Hong Pan, Li Yu Meng, Zhang Hui, Cui Huan-Zhong, Zheng Xin
State Key Laboratory of Animal Nutrition, China Agricultural University, Beijing 100193, China .
Asian-Australas J Anim Sci. 2015 Apr;28(4):573-83. doi: 10.5713/ajas.14.0600.
B-32 is one of a panel of monoclonal anti-idiotypic antibodies to growth hormone (GH) that we developed. To characterize and identify its potential role as a novel growth hormone receptor (GHR) agonist, we determined that B-32 behaved as a typical Ab2β based on a series of enzyme-linked immunosorbent assay assays. The results of fluorescence-activated cell sorting, indirect immunofluorescence and competitive receptor binding assays demonstrated that B-32 specifically binds to the GHR expressed on target cells. Next, we examined the resulting signal transduction pathways triggered by this antibody in primary porcine hepatocytes. We found that B-32 can activate the GHR and Janus kinase (2)/signal transducers and activators of transcription (JAK2/STAT5) signalling pathways. The phosphorylation kinetics of JAK2/STAT5 induced by either GH or B-32 were analysed in dose-response and time course experiments. In addition, B32 could also stimulate porcine hepatocytes to secrete insulin-like growth factors-1. Our work indicates that a monoclonal anti-idiotypic antibody to GH (B-32) can serve as a GHR agonist or GH mimic and has application potential in domestic animal (pig) production.
B-32是我们研发的一组针对生长激素(GH)的单克隆抗独特型抗体之一。为了表征和确定其作为新型生长激素受体(GHR)激动剂的潜在作用,我们通过一系列酶联免疫吸附测定确定B-32表现为典型的Ab2β。荧光激活细胞分选、间接免疫荧光和竞争性受体结合测定的结果表明,B-32特异性结合靶细胞上表达的GHR。接下来,我们研究了该抗体在原代猪肝细胞中触发的信号转导途径。我们发现B-32可以激活GHR和Janus激酶(2)/信号转导子和转录激活子(JAK2/STAT5)信号通路。在剂量反应和时间进程实验中分析了由GH或B-32诱导的JAK2/STAT5的磷酸化动力学。此外,B32还可以刺激猪肝细胞分泌胰岛素样生长因子-1。我们的工作表明,一种针对GH的单克隆抗独特型抗体(B-32)可以作为GHR激动剂或GH模拟物,在家畜(猪)生产中具有应用潜力。