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Single-cell analysis reveals the relevance of foot-and-mouth disease virus persistence to emopamil-binding protein gene expression in host cells

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Abstract

Foot-and-mouth disease virus (FMDV) infects host cells in either an acute or persistent manner. In this study, we examined the relevance of the establishment of FMDV persistence to the expression of the emopamil-binding protein (EBP) gene in 231 individual persistently infected baby hamster kidney (BHK-21) cells after passages 28, 38, and 68 (PI28, PI38, and PI68). At PI28, the stage at which persistent infection of FDMV becomes unstable, the percentage of cells carrying FMDV was 66.7%, while 80.2% of cells were EBP positive. Additionally, in 55.6% of the EBP-positive cells at PI28, EBP expression was upregulated approximately 149.9% compared to uninfected BHK-21 cells. This was the highest expression level among all cell passages measured. Interestingly, in a parallel experiment, the average EBP expression level in the whole cell population at PI28 was only slightly higher (108.2%) than that in uninfected BHK-21 cells. At PI38, 98.7% of the cells were positive for FMDV 3D (an RNA-dependent RNA polymerase enzyme gene), and its maximum expression level observed at this passage. The expression level of EBP in 78.2% of the total cells, however, was reduced significantly. At PI68, 95.8% of the cells were 3D positive, and the expression of both the EBP and 3D genes were at the lowest levels of all the passages. Our studies using single cells yielded data that are otherwise inaccessible a using whole cell population. These results suggest that the establishment of persistent infection by FMDV is a dynamic process that results from the continuous adaptation and coevolution of viruses and cells to reach an equilibrium.

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Acknowledgements

We thank XW and XGD for technical support and helpful discussion during the preparation of the manuscript. This work was funded by the National Natural Sciences Foundation of China (no. 31370185), the National Basic Research Program of China (no. 2011CB504800), and the National Infrastructure of Natural Resources for Science and Technology Program (no. 2011-572) to C. Zheng.

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Authors and Affiliations

  1. State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, China

    Hui Fang, Bing Yuan, Lingling Han, Xiu Xin, Hailong Wang, Congyi Zheng & Chao Shen

  2. Hubei Provincial Cooperative Innovation Center of Industrial Fermentation, Wuhan, China

    Congyi Zheng

  3. China Center for Type Culture Collection, Wuhan University, Wuhan, China

    Congyi Zheng & Chao Shen

  4. Renal Division, Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, 02115, USA

    Fangyan Yu

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  1. Hui Fang
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Contributions

FH and SC conceived the study and designed the experiments. FH, YB, HLL, XX, WHL, and YFY carried out the experimental work. FH, SC, and ZCY analyzed the data and wrote the paper. All authors have read and approved the final manuscript.

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Correspondence to Congyi Zheng or Chao Shen.

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This work does not contain any studies with human participants or animals.

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Handling Editor: Tim Skern.

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Fang, H., Yuan, B., Han, L. et al. Single-cell analysis reveals the relevance of foot-and-mouth disease virus persistence to emopamil-binding protein gene expression in host cells. Arch Virol 162, 3791–3802 (2017). https://doi.org/10.1007/s00705-017-3546-3

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