National Engineering Laboratory for AIDS Vaccine, School of Life Science, Jilin University, Changchun, 130012, China.
Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, School of Life Science, Jilin University, Changchun, 130012, China.
Virol Sin. 2019 Feb;34(1):59-65. doi: 10.1007/s12250-019-00087-3. Epub 2019 Feb 6.
Exosomes are cell-derived vesicles that are secreted by many eukaryotic cells. It has recently attracted attention as vehicles of intercellular communication. Virus-infected cells release exosomes, which contain viral proteins, RNA, and pathogenic molecules. However, the role of exosomes in virus infection process remains unclear and needs to be further investigated. In this study, we aimed to evaluate the effects of exosomes on rabies virus infection. OptiPrep™ density gradient centrifugation was used to isolate exosomes from rabies virus-infected cell culture supernatants. A rabies virus G protein enzyme-linked immunosorbent assay and acetylcholinesterase activity assays were performed to verify the centrifugation fractions. Exosomes were then characterized using transmission electron microscopy and Western blotting. Our results showed that rabies virus infection increased the release of exosomes. Treatment with GW4869 and si-Rab27a, two exosomal secretion inhibitors, inhibited exosome release. Furthermore, the inhibitors reduced the levels of extracellular and intracellular viral RNA. These data indicated that exosomes may participate in the viral infection process. Moreover, our results establish a basis for future research into the roles of exosomes in rabies virus infection and as potential targets for developing new antiviral strategies.
外泌体是由许多真核细胞分泌的细胞来源的囊泡。它最近作为细胞间通讯的载体引起了人们的关注。病毒感染的细胞释放含有病毒蛋白、RNA 和致病分子的外泌体。然而,外泌体在病毒感染过程中的作用尚不清楚,需要进一步研究。在这项研究中,我们旨在评估外泌体对狂犬病病毒感染的影响。使用 OptiPrepTM 密度梯度离心法从狂犬病病毒感染的细胞培养上清液中分离出外泌体。通过狂犬病病毒 G 蛋白酶联免疫吸附测定和乙酰胆碱酯酶活性测定来验证离心部分。然后使用透射电子显微镜和 Western blot 对外泌体进行表征。我们的结果表明,狂犬病病毒感染增加了外泌体的释放。两种外泌体分泌抑制剂 GW4869 和 si-Rab27a 的处理抑制了外泌体的释放。此外,抑制剂降低了细胞外和细胞内病毒 RNA 的水平。这些数据表明外泌体可能参与病毒感染过程。此外,我们的结果为进一步研究外泌体在狂犬病病毒感染中的作用以及作为开发新抗病毒策略的潜在靶点奠定了基础。