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首页> 外文期刊>Journal of interferon and cytokine research: The official journal of the International Society for Interferon and Cytokine Research >A sensitive and versatile bioassay for ligands that signal through receptor clustering.
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A sensitive and versatile bioassay for ligands that signal through receptor clustering.

机译：对通过受体簇发出信号的配体的灵敏且多功能的生物测定。

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摘要

The induced expression of xanthine-guanine phosphoribosyl transferase (XGPRT) by low concentrations (-2 pg/ml) of interferon-alpha (IFN-alpha) or IFN-beta, in the 2fTPGH cell line caused a 50% cytotoxicity when these cells were grown in medium containing 6-thioguanine. We extended the application of this sensitive, reliable, and easy bioassay to other members of the cytokine family. To activate the IFN signaling pathway, we made receptor chimeras, consisting of the IFN type I receptor intracellular and transmembrane domains, fused to either the interleukin-5 (IL-5) receptors or erythropoietin (Epo) receptor extracellular domains as model systems. 2fTGH cells, stably transfected with these receptor chimeras, responded to very low concentrations of IL-5 or Epo (IC50 values of approximately 15 pg and 3 pg/ml, respectively) and thus can be used as a very sensitive bioassay for both ligands. Background activity of IL-5, Epo, tumor necrosis factor (TNF), IL-6, or leptin on cells that did not carry the receptor chimeras was very low. This methodology can in principle be extended to any ligand that acts via clustering of its receptors.
机译：低浓度（-2 pg / ml）的干扰素-α（IFN-alpha）或IFN-β在2fTPGH细胞系中诱导的黄嘌呤-鸟嘌呤磷酸核糖基转移酶（XGPRT）表达导致这些细胞在50％的细胞毒性在含有6-硫鸟嘌呤的培养基中生长。我们将这种灵敏，可靠和简便的生物测定法的应用范围扩展到了细胞因子家族的其他成员。为了激活IFN信号通路，我们制备了由I型受体IFN细胞内和跨膜域组成的受体嵌合体，将其与白细胞介素5（IL-5）受体或促红细胞生成素（Epo）受体胞外域融合作为模型系统。稳定转染了这些受体嵌合体的2fTGH细胞对非常低的IL-5或Epo浓度（分别约为15 pg和3 pg / ml的IC50值）有反应，因此可用作两种配体的非常灵敏的生物测定法。 IL-5，Epo，肿瘤坏死因子（TNF），IL-6或瘦素在不携带受体嵌合体的细胞上的背景活性非常低。原则上，该方法可以扩展到通过其受体聚类起作用的任何配体。

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