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Sci Rep. 2019 Jun 20;9(1):8827. doi: 10.1038/s41598-019-45126-x.

Combating viral contaminants in CHO cells by engineering innate immunity.

Author information

1
Department of Pediatrics, University of California, San Diego, La Jolla, CA, 92093, USA.
2
The Novo Nordisk Foundation Center for Biosustainability at the University of California, San Diego, La Jolla, CA, 92093, USA.
3
Department of Bioengineering, University of California, San Diego, La Jolla, CA, 92093, USA.
4
Bioinformatics and Systems Biology Graduate Program, University of California, San Diego, La Jolla, CA, 92093, USA.
5
Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, 20993, USA.
6
Viral Pathogenesis Section, Laboratory of Immunoregulation, National Institute of Allergy and Infectious Disease, National Institutes of Health, Bethesda, MD, 20892, USA.
7
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark.
8
Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, 20993, USA. Montserrat.puig@fda.hhs.gov.
9
Department of Pediatrics, University of California, San Diego, La Jolla, CA, 92093, USA. nlewisres@ucsd.edu.
10
The Novo Nordisk Foundation Center for Biosustainability at the University of California, San Diego, La Jolla, CA, 92093, USA. nlewisres@ucsd.edu.
11
Department of Bioengineering, University of California, San Diego, La Jolla, CA, 92093, USA. nlewisres@ucsd.edu.

Abstract

Viral contamination in biopharmaceutical manufacturing can lead to shortages in the supply of critical therapeutics. To facilitate the protection of bioprocesses, we explored the basis for the susceptibility of CHO cells to RNA virus infection. Upon infection with certain ssRNA and dsRNA viruses, CHO cells fail to generate a significant interferon (IFN) response. Nonetheless, the downstream machinery for generating IFN responses and its antiviral activity is intact in these cells: treatment of cells with exogenously-added type I IFN or poly I:C prior to infection limited the cytopathic effect from Vesicular stomatitis virus (VSV), Encephalomyocarditis virus (EMCV), and Reovirus-3 virus (Reo-3) in a STAT1-dependent manner. To harness the intrinsic antiviral mechanism, we used RNA-Seq to identify two upstream repressors of STAT1: Gfi1 and Trim24. By knocking out these genes, the engineered CHO cells exhibited activation of cellular immune responses and increased resistance to the RNA viruses tested. Thus, omics-guided engineering of mammalian cell culture can be deployed to increase safety in biotherapeutic protein production among many other biomedical applications.

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