A versatile platform for activity determination of cytokines and growth factors based on the human TSLP (thymic stromal lymphopoietin) receptor
A versatile platform for activity determination of cytokines and growth factors based on the human TSLP (thymic stromal lymphopoietin) receptor
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基于人 TSLP（胸腺基质淋巴细胞生成素）受体的细胞因子和生长因子活性测定的多功能平台
DOI：
10.1016/j.cyto.2018.07.010
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发表时间：
2019
期刊：
影响因子：
3.8
通讯作者：
Wohlmann A
中科院分区：
文献类型：
--
作者：
Marković I;Barthel T;Schirmer M;Gonzälez Delgado A;Wilhelm S;Krause S;Friedrich K;Wohlmann A
细胞因子和生长因子是参与细胞间通讯过程的信号蛋白。它们参与控制许多重要的生理过程，如细胞增殖，基因转录和分化;因此成为基础和应用研究的焦点。其中许多也与人类疾病有关。当作为药理学干预的潜在靶标和结构/功能研究的对象观察时，重要的是以方便和合理的方式测量其生物活性，任选地沿着潜在抑制剂。这样的测试通常是费力的设置和他们的建立是复杂的，因为必须采用有问题的细胞类型和复杂的测定。在这里，我们提出了一个强大的和模块化的活性测定系统，它可以适应几乎所有的配体，通过来自不同家庭的膜受体的二聚化信号。该技术依赖于将特异性受体的配体结合结构域融合到胸腺基质淋巴细胞生成素（TSLP）受体的跨膜和细胞内组分，所述TSLP受体将信号翻译成报告细胞中易于定量的荧光素酶表达。我们表明，各种造血细胞因子受体的激活，受体酪氨酸激酶，以及受体轴承丝氨酸/苏氨酸激酶结构域由各自的配体被忠实地反映后，瞬时和稳定的引入杂交受体和报告基因构建体到小鼠前B细胞系Ba/F3。此外，我们证明了该平台的细胞因子/生长因子受体抑制剂的功能表征的适用性。
Cytokines and growth factors are signaling proteins involved in communication processes between cells. They are involved in the control of numerous essential physiological processes such as cell proliferation, gene transcription and differentiation; therefore being in the focus of basic and applied research. Many of them are also of relevance for human diseases. When observed as potential targets for pharmacological intervention and objects of structure/function studies, it is important to measure their biological activities, optionally along with potential inhibitors, in a convenient and rational manner. Such tests are frequently laborious to set up and their establishment is complicated by the necessity to employ problematic cell types and sophisticated assays. Here we present a robust and modular activity assay system which can be adapted to virtually all ligands that signal through dimerization of membrane receptors from different families. The technique rests on fusing ligand-binding domains of specific receptors to the transmembrane and intracellular components of the thymic stromal lymphopoietin (TSLP) receptor which translates signals into readily quantifiable luciferase expression in reporter cells. We show that the activation of various hematopoietic cytokine receptors, of receptor tyrosine kinases as well as of receptors bearing serine/threonine kinase domains by their respective ligands was faithfully reflected both upon transient and stable introduction of hybrid receptor and reporter gene constructs into the murine pro-B cell line Ba/F3. Moreover, we demonstrate the suitability of this platform for the functional characterization of cytokine/growth factor receptor inhibitors.
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DOI：
--
发表时间：
1997
期刊：
Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS)
影响因子：
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10.1152/ajplung.1994.266.2.l187
发表时间：
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期刊：
The American journal of physiology
影响因子：
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作者：
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作者：
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影响因子：
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作者：
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DOI：
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发表时间：
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期刊：
The Journal of biological chemistry
影响因子：
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作者：
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通讯作者：
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