Zhang Qian, Zeng Lei-Ping, Zhou Peng, Irving Aaron T, Li Shang, Shi Zheng-Li, Wang Lin-Fa
Key Laboratory of Special Pathogens, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China.
Programme in Emerging Infectious Diseases, Duke-National University of Singapore Medical School, Singapore, Singapore.
PLoS One. 2017 Aug 9;12(8):e0182866. doi: 10.1371/journal.pone.0182866. eCollection 2017.
Bats are important reservoirs of many viruses, which are capable of infecting the host without inducing obvious clinical diseases. Interferon and the downstream interferon regulated genes (IRGs) are known to act as the first line of defense against viral infections. Little is known about the transcriptional profile of genes being induced by interferon in bats and their role in controlling virus infection. In this study, we constructed IFNAR2 knockout bat cell lines using CRISPR technology and further characterized gene expression profiles induced by the most abundant IFN-α (IFN-α3). Firstly, we demonstrated that the CRISPR/Cas9 system is applicable for bat cells as this represents the first CRIPSR knockout cell line for bats. Our results showed the pleiotropic effect of IFN-α3 on the bat kidney cell line, PaKiT03. As expected, we confirmed that IFNAR2 is indispensable for IFN-a signaling pathway and plays an important role in antiviral immunity. Unexpectedly, we also identified novel IFNAR2-dependent IRGs which are enriched in pathways related to cancer. To our knowledge, this seems to be bat-specific as no such observation has been reported for other mammalian species. This study expands our knowledge about bat immunology and the cell line established can provide a powerful tool for future study into virus-bat interaction and cancer biology.
蝙蝠是许多病毒的重要宿主，这些病毒能够在不引发明显临床疾病的情况下感染宿主。干扰素及下游干扰素调节基因（IRGs）被认为是抵御病毒感染的第一道防线。关于蝙蝠体内由干扰素诱导的基因转录谱及其在控制病毒感染中的作用，我们所知甚少。在本研究中，我们利用CRISPR技术构建了IFNAR2基因敲除的蝙蝠细胞系，并进一步对最丰富的IFN-α（IFN-α3）诱导的基因表达谱进行了表征。首先，我们证明CRISPR/Cas9系统适用于蝙蝠细胞，因为这是首个针对蝙蝠的CRIPSR基因敲除细胞系。我们的结果显示了IFN-α3对蝙蝠肾细胞系PaKiT03的多效性作用。正如预期的那样，我们证实IFNAR2对于IFN-α信号通路不可或缺，且在抗病毒免疫中发挥重要作用。出乎意料的是，我们还鉴定出了新的依赖IFNAR2的IRGs，这些基因在与癌症相关的通路中富集。据我们所知，这似乎是蝙蝠特有的现象，因为其他哺乳动物物种尚未有此类报道。本研究扩展了我们对蝙蝠免疫学的认识，所建立的细胞系可为未来研究病毒与蝙蝠的相互作用以及癌症生物学提供有力工具。
