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使用荧光金属纳米探针直接观察 T 淋巴细胞表面的趋化因子受体 5。

Direct observation to chemokine receptor 5 on T-lymphocyte cell surface using fluorescent metal nanoprobes.

机构信息

Center for Fluorescence Spectroscopy, Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, 725 West Lombard Street, Baltimore, MD 21201, USA.

出版信息

Biochem Biophys Res Commun. 2010 Sep 10;400(1):111-6. doi: 10.1016/j.bbrc.2010.08.020. Epub 2010 Aug 10.

Abstract

Chemokine receptor 5 (CCR5) is a cell surface protein required for HIV-1 infection. It is important to detect the amount and observe the spatial distribution of the CCR5 receptors on the cell surfaces. In this report, we describes the metal nanoparticles which were specially designed as molecular fluorescent probes for imaging of CCR5 receptors on the T-lymphocytic PM1 cell surfaces. These CCR5 monoclonal antibodies (mAbs) metal complexes were prepared by labeling mAbs with Alexa Fluor 680 followed by covalent binding the labeled mAbs on the 20 nm silver nanoparticles. Compared with the labeled mAbs without metal, the mAb-metal complexes were found to display enhanced emission intensity and shortened lifetime due to interactions between fluorophores and metal. The mAb-metal complexes were incubated with the PM1 cell lines. The confocal fluorescent intensity and lifetime cell images were recorded on single cells. It was observed that the mAb-metal complexes could be clearly distinguished from the cellular autofluorescence. By analyzing a pool of cell images, we observed that most CCR5 receptors appeared as clusters on the cell surfaces. The fluorophore-metal complexes developed in this report are generally useful for detection of cell surface receptors and provide a new class of probe to study the interaction between the CCR5 receptors with viral gp120 during HIV infections.

摘要

趋化因子受体 5(CCR5)是 HIV-1 感染所必需的细胞表面蛋白。检测细胞表面 CCR5 受体的数量并观察其空间分布非常重要。在本报告中,我们描述了专门设计的金属纳米粒子作为用于 PM1 淋巴细胞表面 CCR5 受体成像的分子荧光探针。这些 CCR5 单克隆抗体(mAbs)金属复合物是通过用 Alexa Fluor 680 标记 mAbs 然后将标记的 mAbs 共价结合到 20nm 的银纳米粒子上来制备的。与没有金属的标记 mAbs 相比,由于荧光团和金属之间的相互作用,mAb-金属复合物显示出增强的发射强度和缩短的寿命。将 mAb-金属复合物与 PM1 细胞系一起孵育。在单细胞上记录共聚焦荧光强度和寿命细胞图像。观察到 mAb-金属复合物可以与细胞自发荧光明显区分开来。通过分析一组细胞图像,我们观察到大多数 CCR5 受体在细胞表面呈现为簇。本报告中开发的荧光团-金属复合物通常可用于检测细胞表面受体,并为研究 HIV 感染期间 CCR5 受体与病毒 gp120 之间的相互作用提供了一类新的探针。

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