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Antigen-Responsive Regulation of Cell Motility and Migration via the Signalobodies Based on c-Fms and c-MpI

机译:基于c-Fms和c-MpI的信号体对细胞运动和迁移的抗原响应调节

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Since cell migration plays critical roles in development and homeostasis of the body, artificial control of cell migration would be promising for the treatment of various diseases related to migration. To this end, we previously developed single-chain Fv (scFv)lreceptor chimeras, named signalobodies, which can control cell fates via a specific antigen that is different from natural cytokines. Although a conventional chemotaxis chamber assay revealed that several signalobodies based on receptor tyrosine kinases transduced antigen-dependent migration signals, we have never performed direct observation of the cells to obtain more information on overall properties of cell motility and migration. In this study, we utilized murine pro-B BalF3 cells expressing either a scFv-Fms or scFv-Mpl signalobody, and compared their migratory characteristics. We employed a lipid-polyethylene glycol conjugate to softly immobilize the suspension cells on a slide, which facilitated direct observation of chemokinetic activity of the cells. Consequently, both cells markedly exhibited chemokinesis in response to a specific antigen. In addition, the cells were subjected to a stable antigen-concentration gradient to observe horizontal directional cell migration in real time. The results showed that the cells expressing scFv-Fms underwent directional migration toward a positive antigen-concentration gradient. Taken together, we successfully demonstrated antigen-responsive regulation of cell motility and migration via the signalobodies.
机译:由于细胞迁移在人体的发育和体内平衡中起着关键作用,因此人工控制细胞迁移对于治疗与迁移有关的各种疾病将是有希望的。为此,我们先前开发了单链Fv(scFv)受体嵌合体,称为信号体,它可以通过不同于天然细胞因子的特定抗原控制细胞命运。尽管常规的趋化性室测定法揭示了几种基于受体酪氨酸激酶的信号体可转导抗原依赖性迁移信号,但我们从未对细胞进行直接观察以获取有关细胞运动和迁移的整体特性的更多信息。在这项研究中,我们利用了表达scFv-Fms或scFv-Mpl信号体的鼠pro-B BalF3细胞,并比较了它们的迁移特性。我们采用脂质-聚乙二醇共轭物将悬浮细胞轻轻固定在载玻片上,这有助于直接观察细胞的化学动力学活性。因此,两种细胞都对特定抗原有明显的趋化作用。另外,使细胞经受稳定的抗原浓度梯度,以实时观察水平方向的细胞迁移。结果表明表达scFv-Fms的细胞经历了向正抗原浓度梯度的定向迁移。两者合计,我们成功地证明了通过信号抗体对细胞运动性和迁移的抗原响应调节。

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