In vitro and in vivo characterization of recombinant Ebola viruses expressing enhanced green fluorescent protein
In vitro and in vivo characterization of recombinant Ebola viruses expressing enhanced green fluorescent protein
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DOI：
10.1086/520590
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发表时间：
2007-11-15
影响因子：
6.4
通讯作者：
Feldmann, Heinz
中科院分区：
文献类型：
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作者：
Ebihara, Hideki;Theriault, Steven;Feldmann, Heinz
为了促进对扎伊尔埃博拉病毒（ZEBOV）感染的发病机理的分子方面的理解，我们从插入在病毒基因组中的不同位置的另外的转录单位产生了表达增强型绿色荧光蛋白（eGFP）的2种不同的重组病毒。这些病毒显示出与野生型ZEBOV（wt-ZEBOV）相似的体外表型，并且在多次传代中是稳定的。用表达eGFP的病毒之一感染恒河猴仅产生轻度疾病，在该动物模型中显示出显著的减毒。然而，在缺乏信号转导子和转录激活子1的小鼠中，表达eGFP的两种病毒引起的致死性疾病与由wt-ZEBOV引起的相比是中度减弱的。在小鼠中，可以通过使用流式细胞术检测组织中的eGFP阳性细胞来容易地跟踪病毒复制。这些发现表明，外源基因的掺入将在体内减弱ZEBOV，但这些病毒仍然具有体外和体内研究应用的潜力。
To facilitate an understanding of the molecular aspects of the pathogenesis of Zaire ebolavirus (ZEBOV) infection, we generated 2 different recombinant viruses expressing enhanced green fluorescent protein (eGFP) from additional transcription units inserted at different positions in the virus genome. These viruses showed in vitro phenotypes similar to that of wild-type ZEBOV (wt-ZEBOV) and were stable over multiple passages. Infection with one of the viruses expressing eGFP produced only mild disease in rhesus macaques, demonstrating a marked attenuation in this animal model. However, in mice lacking signal transducer and activator of transcription 1, both viruses expressing eGFP caused lethal cases of disease that were moderately attenuated, compared with that caused by wt-ZEBOV. In mice, viral replication could be easily tracked by the detection of eGFP-positive cells in tissues, by use of flow cytometry. These findings demonstrate that the incorporation of a foreign gene will attenuate ZEBOV in vivo but that these viruses still have potential for in vitro and in vivo research applications.