Visualization of chemokine binding sites on human brain microvessels
Visualization of chemokine binding sites on human brain microvessels
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DOI：
10.1083/jcb.145.2.403
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发表时间：
1999-04-19
影响因子：
7.8
通讯作者：
Pachter, JS
中科院分区：
文献类型：
--
作者：
Andjelkovic, AV;Spencer, DD;Pachter, JS
趋化因子单核细胞趋化蛋白-1（MCP-1）和巨噬细胞炎性蛋白-1 α\（MIP-1 α）有助于在中枢神经系统炎症期间将白细胞引导至脑和脊髓内的特定部位。然而，目前还不清楚这些趋化因子如何跨越血管屏障发挥作用，这引起了人们的猜测，即可能需要与内皮细胞相互作用。因此，进行实验以确定这些趋化因子的结合结构域是否存在于脑微血管的外表面沿着，这是一种可能将趋化因子信号从脑传递到血液的特征。使用生物素化的趋化因子结合分析与共聚焦显微镜和三维图像重建，MCP-1和MIP-α周围的人脑微血管的空间分辨的结合位点被首次揭示。标记的MCP-1和MIP-1 α的结合可以被未标记的同源而非异源趋化因子抑制，并且不依赖于内皮下基质中硫酸乙酰肝素、层粘连蛋白或胶原蛋白的存在。这是MCP-1和MIP-1 α在微血管的实质表面上特异性和单独结合域的第一个证据，并突出了趋化因子与微血管成分的特异性相互作用影响中枢神经系统炎症的程度和过程的前景。
The chemokines monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-1 alpha\ (MIP-1 alpha) aid in directing leukocytes to specific locales within the brain and spinal cord during central nervous system inflammation. However, it remains unclear how these chemokines exert their actions across a vascular barrier, raising speculation that interaction with endothelial cells might be required. Therefore, experiments were performed to determine whether binding domains for these chemokines exist along the outer surface of brain microvessels, a feature that could potentially relay chemokine signals from brain to blood. Using a biotinylated chemokine binding assay with confocal microscopy and three-dimensional image reconstruction, spatially resolved binding sites for MCP-1 and MIP-alpha around human brain microvessels were revealed for the first time. Binding of labeled MCP-1 and MIP-1 alpha could be inhibited by unlabeled homologous but not heterologous chemokine, and was independent of the presence of heparan sulfate, laminin, or collagen in the subendothelial matrix. This is the first evidence of specific and separate binding domains for MCP-1 and MIP-1 alpha, on the parenchymal surface of microvessels, and highlights the prospect that specific interactions of chemokines with microvascular elements influence the extent and course of central nervous system inflammation.