Zeng Yu-Jing, Hsu Min-Kung, Tsai Chiao-An, Chu Chun-Yen, Wu Hsing-Chieh, Wang Hsian-Yu
International Program in Animal Vaccine Technology, International College, National Pingtung University of Science and Technology, Pingtung 91201, Taiwan.
Research Center for Tumor Medical Science, China Medical University, Taichung 40402, Taiwan.
Vaccines (Basel). 2021 Jun 4;9(6):601. doi: 10.3390/vaccines9060601.
During industrial-scale production of viruses for vaccine manufacturing, anti-viral response of host cells can dampen maximal viral antigen yield. In addition to interferon responses, many other cellular responses, such as the AMPK signaling pathway or senescence-like response may inhibit or slow down virus amplification in the cell culture system. In this study, we first performed a Gene Set Enrichment Analysis of the whole-genome mRNA transcriptome and found a senescence-like cellular response in BHK-21 cells when infected with bovine ephemeral fever virus (BEFV). To demonstrate that this senescence-like state may reduce virus growth, BHK-21 subclones showing varying degrees of a senescence-like state were infected with BEFV. The results showed that the BHK-21 subclones showing high senescence staining could inhibit BEFV replication while low senescence-staining subclones are permissive to virus replication. Using a different approach, a senescence-like state was induced in BHK-21 using a small molecule, camptothecin (CPT), and BEFV susceptibility were examined. The results showed that CPT-treated BHK-21 is more resistant to virus infection. Overall, these results indicate that a senescence-like response may be at play in BHK-21 upon virus infection. Furthermore, cell clone selection and modulating treatments using small molecules may be tools in countering anti-viral responses.
在用于疫苗生产的病毒进行工业规模生产期间，宿主细胞的抗病毒反应会抑制病毒抗原的最大产量。除了干扰素反应外，许多其他细胞反应，如AMPK信号通路或类似衰老的反应，可能会抑制或减缓细胞培养系统中的病毒扩增。在本研究中，我们首先对全基因组mRNA转录组进行了基因集富集分析，发现感染牛暂时热病毒（BEFV）时，BHK-21细胞中存在类似衰老的细胞反应。为了证明这种类似衰老的状态可能会降低病毒生长，用BEFV感染了表现出不同程度类似衰老状态的BHK-21亚克隆。结果表明，高衰老染色的BHK-21亚克隆可以抑制BEFV复制，而低衰老染色的亚克隆则允许病毒复制。采用不同的方法，用小分子喜树碱（CPT）在BHK-21中诱导出类似衰老的状态，并检测BEFV的易感性。结果表明，CPT处理的BHK-21对病毒感染更具抗性。总体而言，这些结果表明，病毒感染后BHK-21中可能存在类似衰老的反应。此外，细胞克隆选择和使用小分子的调节处理可能是对抗抗病毒反应的工具。
