单克隆拮抗剂抗体对降钙素基因相关肽受体功能和转运的影响。
Effects of monoclonal antagonist antibodies on calcitonin gene-related peptide receptor function and trafficking.
机构信息
Department of Cytometry Sciences, Amgen Research, 360 Binney Street, Cambridge, MA, 02142, USA.
Department of Neuroscience, Amgen Research, 360 Binney Street, Cambridge, MA, 02142, USA.
出版信息
J Headache Pain. 2019 Apr 30;20(1):44. doi: 10.1186/s10194-019-0992-1.
BACKGROUND
Monoclonal antibodies against calcitonin gene-related peptide (CGRP) or its receptor are efficacious for the prevention of migraine headaches. The downstream molecular mechanisms following ligand-receptor blockade by which these antibodies prevent CGRP signaling through CGRP receptors have not been demonstrated.
METHODS
Here we produced tool monoclonal functional antagonist antibodies against CGRP and its canonical receptor and developed a novel cellular model using fluorogen-activated protein technology that allows detection of CGRP receptor internalization by flow cytometry and, for an extended time course, visualization by confocal microscopy.
RESULTS
Using this cell model we showed that these antagonist antibodies block both CGRP-induced cAMP signaling and CGRP receptor internalization. At least 10-fold higher concentrations of either antibody are necessary to block CGRP receptor internalization compared with cAMP accumulation in our cell model.
CONCLUSION
These data reinforce our understanding of how monoclonal functional antagonist antibodies interfere with CGRP signaling.
背景
针对降钙素基因相关肽（CGRP）或其受体的单克隆抗体可有效预防偏头痛。但这些抗体通过配体-受体阻断来阻止 CGRP 信号通过 CGRP 受体的下游分子机制尚未得到证实。
方法
我们在此制备了针对 CGRP 及其经典受体的工具性单克隆功能拮抗抗体，并开发了一种新的细胞模型，该模型使用荧光素酶激活蛋白技术，可通过流式细胞术检测 CGRP 受体内化，并通过共聚焦显微镜进行延长时间过程的可视化。
结果
使用该细胞模型，我们表明这些拮抗剂抗体可阻断 CGRP 诱导的 cAMP 信号和 CGRP 受体内化。与我们细胞模型中 cAMP 积累相比，阻断 CGRP 受体内化所需的单克隆抗体浓度至少高 10 倍。
结论
这些数据强化了我们对单克隆功能拮抗抗体如何干扰 CGRP 信号的理解。
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