Investigation of Influenza Virus Polymerase Activity in Pig Cells
Investigation of Influenza Virus Polymerase Activity in Pig Cells
复制标题
DOI：
10.1128/jvi.01633-12
复制
发表时间：
2013-01-01
影响因子：
5.4
通讯作者：
Barclay, Wendy S.
中科院分区：
文献类型：
--
作者：
Moncorge, Olivier;Long, Jason S.;Barclay, Wendy S.
在猪中经常检测到具有禽类、人类和/或猪基因组片段组合的重配流感病毒。因此，猪被指责为流感病毒的“混合容器”。这意味着猪细胞支持禽流感病毒的转录和复制，而人类细胞中大多数禽流感病毒聚合酶的活性有限。尽管多年来通过使用小基因组测定在人类和禽类细胞中研究流感病毒聚合酶活性，但在猪细胞中的类似研究尚未见报道。我们开发了第一个针对猪细胞的小基因组测定，并比较了猪、人类和禽细胞中禽流感病毒或人流感病毒来源的聚合酶的活性。我们还在猪细胞中研究了一些已知的哺乳动物宿主范围决定因素（例如 PB2 突变 E627K、D701N、G590S/Q591R 和 T271A）增强人类细胞中流感病毒聚合酶活性的后果。本研究中使用的两种典型禽流感病毒聚合酶在猪细胞中的活性较差，与在人类细胞中观察到的情况相似，并且使禽流感病毒聚合酶适应人类细胞的突变也增加了猪细胞中的活性。相比之下，在鸟类细胞中观察到不同的模式。最后，测试了高致病性禽流感病毒 H5N1 聚合酶活性，因为据报道该亚型在猪体内的复制能力很差。 H5N1 聚合酶在猪细胞中活跃，表明其他屏障限制这些病毒在猪中流行。
Reassortant influenza viruses with combinations of avian, human, and/or swine genomic segments have been detected frequently in pigs. As a consequence, pigs have been accused of being a "mixing vessel" for influenza viruses. This implies that pig cells support transcription and replication of avian influenza viruses, in contrast to human cells, in which most avian influenza virus polymerases display limited activity. Although influenza virus polymerase activity has been studied in human and avian cells for many years by use of a minigenome assay, similar investigations in pig cells have not been reported. We developed the first minigenome assay for pig cells and compared the activities of polymerases of avian or human influenza virus origin in pig, human, and avian cells. We also investigated in pig cells the consequences of some known mammalian host range determinants that enhance influenza virus polymerase activity in human cells, such as PB2 mutations E627K, D701N, G590S/Q591R, and T271A. The two typical avian influenza virus polymerases used in this study were poorly active in pig cells, similar to what is seen in human cells, and mutations that adapt the avian influenza virus polymerase for human cells also increased activity in pig cells. In contrast, a different pattern was observed in avian cells. Finally, highly pathogenic avian influenza virus H5N1 polymerase activity was tested because this subtype has been reported to replicate only poorly in pigs. H5N1 polymerase was active in swine cells, suggesting that other barriers restrict these viruses from becoming endemic in pigs.