Role of Phosphatidylserine-Derived Negative Surface Charges in the Recognition and Uptake of Intravenously Injected B16BL6-Derived Exosomes by Macrophages
Role of Phosphatidylserine-Derived Negative Surface Charges in the Recognition and Uptake of Intravenously Injected B16BL6-Derived Exosomes by Macrophages
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DOI：
10.1016/j.xphs.2016.07.022
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发表时间：
2017-01-01
影响因子：
3.8
通讯作者：
Takakura, Yoshinobu
中科院分区：
文献类型：
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作者：
Matsumoto, Akihiro;Takahashi, Yuki;Takakura, Yoshinobu
外来体是细胞来源的细胞外囊泡，其充当细胞间递送载体。我们先前的研究表明，肝脏中的巨噬细胞有助于静脉内施用的B16 BL 6衍生的外泌体从小鼠体循环中的快速清除。磷脂酰丝氨酸（PS）可能负责这种清除，因为它暴露在外泌体的表面上，并被巨噬细胞识别。在本研究中，研究了暴露于外泌体膜上的PS在巨噬细胞摄取B16 BL 6衍生的外泌体中的作用。带负电荷的PS或磷脂酰甘油负载的脂质体抑制PKH 67标记的外泌体的巨噬细胞的细胞摄取，而含磷脂酰胆碱的脂质体不影响摄取。随后，对于体内分析，使用包含外泌体亲性蛋白质lactadherin的外泌体融合蛋白，用Gaussia荧光素酶、报告蛋白或（3-I-125-碘苯甲酰基）降生物素酰胺标记外泌体。预先注射含有PS或磷脂酰甘油的脂质体后，静脉注射到小鼠体内后，Gaussia β-淀粉酶标记的外泌体的血液清除率显著延迟。此外，（3-I-125-碘苯甲酰基）降生物素酰胺标记的外泌体在肝脏中的蓄积通过预注射含PS的脂质体而降低。这些结果表明外泌体膜中PS的负电荷参与巨噬细胞对静脉注射的外泌体的识别和清除。(C)2016年美国药学协会（R）。爱思唯尔公司出版All rights reserved.
Exosomes are cell-derived extracellular vesicles that function as intercellular delivery carriers. Our previous study demonstrated that macrophages in the liver contributed to the rapid clearance of intravenously administered B16BL6-derived exosomes from the systemic circulation in mice. Phosphatidylserine (PS) may be responsible for this clearance because it is exposed on the surface of exosomes and is recognized by macrophages. In this study, the role of PS exposed on the membranes of exosomes in the uptake of B16BL6-derived exosomes by macrophages was investigated. Negatively charged PS-or phosphatidylglycerol-loaded liposomes suppressed the cellular uptake of PKH67-labeled exosomes by macrophages, whereas phosphatidylcholine-containing liposome did not affect uptake. Subsequently, for the in vivo analysis, exosomes were labeled with Gaussia luciferase, a reporter protein, or (3-I-125-iodobenzoyl) norbiotinamide using exosome-tropic fusion proteins comprising the exosome-tropic protein lactadherin. The blood clearance of Gaussia luciferase-labeled exosomes after intravenous injection into mice was significantly delayed by the preinjection of PS-or phosphatidylglycerol-containing liposomes. Moreover, the accumulation of (3-I-125-iodobenzoyl)norbiotinamide-labeled exosomes in the liver was decreased by the preinjection of PS-containing liposomes. These results indicate that the negative charge of PS in exosomal membranes is involved in the recognition and clearance of intravenously injected exosomes by macrophages. (C) 2016 American Pharmacists Association (R). Published by Elsevier Inc. All rights reserved.