Study of the transcytosis of an anti-transferrin receptor antibody with a Fab' cargo across the blood-brain barrier in mice
Study of the transcytosis of an anti-transferrin receptor antibody with a Fab' cargo across the blood-brain barrier in mice
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DOI：
10.1016/j.ejps.2013.05.027
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发表时间：
2013-07-16
影响因子：
4.6
通讯作者：
Vilaplana, Jordi
中科院分区：
文献类型：
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作者：
Manich, Gemma;Cabezon, Itsaso;Vilaplana, Jordi
一种用于转运抗肿瘤活性物质穿过血脑屏障（BBB）的策略是使用脑毛细血管内皮细胞中的受体介导的跨细胞转运系统将物质连接至能够穿过BBB的分子。转铁蛋白受体（TfR）与这些细胞中的转胞吞过程相关，并且针对小鼠TfR的8D 3抗体能够诱导受体应答。在这项工作中，研究了803携带分子穿过BBB的潜力。将该抗体（未标记或FITC标记）静脉内给予ICR-CD 1小鼠。还向ICR-CD 1小鼠施用由8D 3抗体（载体）和Fab'片段（模拟货物）组成的免疫复合物（IC），在某些情况下直接用FITC荧光标记。在不同的时间点的再循环，IC组件进行了研究，使用免疫组织化学程序和荧光共振能量转移（FRET）。结果表明，8D 3单独或与Fab'片段的IC进入内皮细胞。FRET分析表明，它们的荧光信号在内皮细胞内的共定位随时间减少，表明IC可以被加工，并且Fab'片段可能与8D 3分离。然而，无论是单独的8D 3还是IC组件都没有穿过BBB。(C)2013爱思唯尔有限公司版权所有。
One strategy used to transport pharmacologically active substances across the blood-brain barrier (BBB) is to link the substance to a molecule capable of crossing the BBB using a receptor-mediated transcellular transport system in brain capillary endothelial cells. The transferrin receptor (TfR) is related to a transcytosis process in these cells, and the 8D3 antibody, directed against mouse TfR, is able to induce a receptor response. In this work, the potential of 803 to carry molecules across the BBB was investigated. This antibody (either unlabeled or FITC-labeled) was intravenously administered to ICR-CD1 mice. Immuno-complexes (ICs) consisting of 8D3 antibody (carrier) and Fab' fragments (simulated cargo), in some cases directly fluorolabeled with FITC, were also administered to ICR-CD1 mice. At different time-points of recirculation, the IC components were studied using immunohistochemical procedures and fluorescence resonance energy transfer (FRET). The results suggested that 8D3 alone or in an IC with Fab' fragments entered the endothelial cells. FRET analysis indicated that the colocalization of their fluorescent signals inside the endothelial cells decreased with time, indicating that ICs can be processed and Fab' fragments probably separated from 8D3. However, neither 8D3 alone nor the IC components crossed the BBB. (C) 2013 Elsevier B.V. All rights reserved.