Macrophage-dependent clearance of systemically administered B16BL6-derived exosomes from the blood circulation in mice.
Macrophage-dependent clearance of systemically administered B16BL6-derived exosomes from the blood circulation in mice.
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DOI：
10.3402/jev.v4.26238
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发表时间：
2015
影响因子：
16
通讯作者：
Takakura Y
中科院分区：
文献类型：
--
作者：
Imai T;Takahashi Y;Nishikawa M;Kato K;Morishita M;Yamashita T;Matsumoto A;Charoenviriyakul C;Takakura Y
先前使用标记有gLuc - 乳黏附素（gLuc - LA，一种高斯荧光素酶（一种报告蛋白）和乳黏附素（一种亲外泌体蛋白）的融合蛋白）的B16BL6来源的外泌体的研究表明，在小鼠静脉注射后，外泌体迅速从体循环中消失。在本研究中，对静脉注射的B16BL6外泌体快速清除的机制进行了研究。用编码gLuc - LA的质粒DNA转染B16BL6细胞后，从其细胞上清液中获得gLuc - LA标记的外泌体。当外泌体在血清中孵育时，标记稳定。通过使用用亲脂性荧光染料PKH26标记的B16BL6外泌体，证明PKH26标记的B16BL6外泌体被肝脏和脾脏中的巨噬细胞摄取，但不被肺中的巨噬细胞摄取，而PKH26标记的外泌体被肺中的内皮细胞摄取。随后，将gLuc - LA标记的B16BL6外泌体注射到通过注射含氯膦酸盐脂质体制备的巨噬细胞耗竭小鼠体内。静脉注射的B16BL6外泌体从血液循环中的清除在巨噬细胞耗竭小鼠中比在未处理小鼠中慢得多。这些结果表明巨噬细胞在静脉注射的B16BL6外泌体从体循环的清除中起重要作用。
Previous studies using B16BL6-derived exosomes labelled with gLuc–lactadherin (gLuc-LA), a fusion protein of Gaussia luciferase (a reporter protein) and lactadherin (an exosome-tropic protein), showed that the exosomes quickly disappeared from the systemic circulation after intravenous injection in mice. In the present study, the mechanism of rapid clearance of intravenously injected B16BL6 exosomes was investigated. gLuc-LA-labelled exosomes were obtained from supernatant of B16BL6 cells after transfection with a plasmid DNA encoding gLuc-LA. Labelling was stable when the exosomes were incubated in serum. By using B16BL6 exosomes labelled with PKH26, a lipophilic fluorescent dye, it was demonstrated that PKH26-labelled B16BL6 exosomes were taken up by macrophages in the liver and spleen but not in the lung, while PKH26-labelled exosomes were taken up by the endothelial cells in the lung. Subsequently, gLuc-LA-labelled B16BL6 exosomes were injected into macrophage-depleted mice prepared by injection with clodronate-containing liposomes. The clearance of the intravenously injected B16BL6 exosomes from the blood circulation was much slower in macrophage-depleted mice than that in untreated mice. These results indicate that macrophages play important roles in the clearance of intravenously injected B16BL6 exosomes from the systemic circulation.