Construction of a Fluorescein-Responsive Chimeric Receptor With Strict Ligand Dependency
Construction of a Fluorescein-Responsive Chimeric Receptor With Strict Ligand Dependency
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DOI：
10.1002/bit.21961
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发表时间：
2008-12-01
影响因子：
3.8
通讯作者：
Nagamune, Teruyuki
中科院分区：
文献类型：
--
作者：
Liu, Wenhai;Kawahara, Masahiro;Nagamune, Teruyuki
由于许多细胞功能是由细胞因子受体超家族的成员，细胞因子受体系统的人工模拟将是有吸引力的细胞工程。我们先前表明，抗体/细胞因子受体嵌合体可以响应于非天然配体（如荧光素缀合的BSA）而抑制生长信号。然而，在没有抗原的情况下观察到相当大的细胞增殖背景。因此，我们重新设计了嵌合受体的结构域的不同组合，含有抗荧光素单链抗体（ScFv），细胞外D1/D2以及红细胞生成素受体（EpoR）的跨膜结构域，和糖蛋白130（gp 130）的细胞内结构域，以获得严格的荧光素依赖性嵌合受体。当用编码单个嵌合受体的逆转录病毒载体转导白细胞介素-3依赖性Ba/F3细胞时，具有D1和D2结构域或不具有任何EpoR胞外结构域的嵌合体获得严格的配体依赖性开/关调节。
Since many cell functions are regulated by members of the cytokine receptor superfamily, the artificial mimicry of the cytokine receptor system would be attractive for cellular engineering. We previously showed that an antibody/cytokine receptor chimera can transduce a growth signal in response to non-natural ligands, such as fluorescein-conjugated BSA. However, considerable background of cell proliferation was observed without antigen. Therefore, we redesigned chimeric receptor constructs with different combinations of domains containing anti-fluorescein single chain Fv (ScFv), extracellular D1/D2 as well as transmembrane domains of erythropoietin receptor (EpoR), and the intracellular domain of glycoprotein 130 (gp130), to obtain strictly fluorescein-dependent chimeric receptors. When interleukin-3-dependent Ba/F3 cells were transduced with retroviral vectors encoding individual chimeric receptors, the chimeras either with both D I and D2 domains or without any EpoR extracellular domain attained a strict ligand-dependent ON/OFF regulation.