Enhanced isolation of SARS-CoV-2 by TMPRSS2-expressing cells

doi:10.1073/pnas.2002589117

. 2020 Mar 31;117(13):7001-7003.

doi: 10.1073/pnas.2002589117. Epub 2020 Mar 12.

Enhanced isolation of SARS-CoV-2 by TMPRSS2-expressing cells

Shutoku Matsuyama¹, Naganori Nao², Kazuya Shirato², Miyuki Kawase², Shinji Saito³, Ikuyo Takayama³, Noriyo Nagata⁴, Tsuyoshi Sekizuka⁵, Hiroshi Katoh², Fumihiro Kato², Masafumi Sakata², Maino Tahara², Satoshi Kutsuna⁶, Norio Ohmagari⁶, Makoto Kuroda⁵, Tadaki Suzuki⁴, Tsutomu Kageyama³, Makoto Takeda¹

Affiliations

¹ Department of Virology 3, National Institute of Infectious Diseases, Tokyo 162-8640, Japan; matuyama@nih.go.jp mtakeda@nih.go.jp.
² Department of Virology 3, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.
³ Influenza Virus Research Center, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.
⁴ Department of Pathology, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.
⁵ Pathogen Genomics Center, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.
⁶ Disease Control and Prevention Center, National Center for Global Health and Medicine, Tokyo 162-8655, Japan.

PMID: 32165541
PMCID: PMC7132130
DOI: 10.1073/pnas.2002589117

Enhanced isolation of SARS-CoV-2 by TMPRSS2-expressing cells

Shutoku Matsuyama et al. Proc Natl Acad Sci U S A. 2020.

. 2020 Mar 31;117(13):7001-7003.

doi: 10.1073/pnas.2002589117. Epub 2020 Mar 12.

Authors

Affiliations

¹ Department of Virology 3, National Institute of Infectious Diseases, Tokyo 162-8640, Japan; matuyama@nih.go.jp mtakeda@nih.go.jp.
² Department of Virology 3, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.
³ Influenza Virus Research Center, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.
⁴ Department of Pathology, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.
⁵ Pathogen Genomics Center, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.
⁶ Disease Control and Prevention Center, National Center for Global Health and Medicine, Tokyo 162-8655, Japan.

PMID: 32165541
PMCID: PMC7132130
DOI: 10.1073/pnas.2002589117

Abstract

A novel betacoronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which caused a large respiratory outbreak in Wuhan, China in December 2019, is currently spreading across many countries globally. Here, we show that a TMPRSS2-expressing VeroE6 cell line is highly susceptible to SARS-CoV-2 infection, making it useful for isolating and propagating SARS-CoV-2. Our results reveal that, in common with SARS- and Middle East respiratory syndrome-CoV, SARS-CoV-2 infection is enhanced by TMPRSS2.

Keywords: SARS-CoV-2; TMPRSS2; VeroE6; coronavirus; outbreak.

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Conflict of interest statement

The authors declare no competing interest.

Figures

**Fig. 1.**
(A) Expression of TMPRSS2 in total cellular RNA (0.2 µg) of indicated cells was compared with that in human lung RNA (catalog no. 636524; Clontech) by quantitative real-time PCR. ND, not detectable. (B) SARS-CoV-2–infected VeroE6/TMPRSS2 cells. Cell rounding (black arrows) and syncytium formation (white arrows) (C). Electron micrograph showing isolated virus particles with negative staining. (Scale bar, 200 nm.) (D) Viral RNA multiplication in various cells at 48 h postinoculation with the viral specimen, as determined by real-time RT-PCR using E and N primer/probe sets (9). Cq, quantitation cycle. (E) Real-time RT-PCR amplification plot using the E primer/probe set, corresponding to the data in C. RFU, relative fluorescence units. (F) Comparison of cell susceptibility to the isolated virus, detected with a patient’s serum and Alexa 488-conjugated goat anti-human IgG. Nuclei were stained with DAPI.

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Comment in

Possible use of the mucolytic drug, bromhexine hydrochloride, as a prophylactic agent against SARS-CoV-2 infection based on its action on the Transmembrane Serine Protease 2.
Habtemariam S, Nabavi SF, Ghavami S, Cismaru CA, Berindan-Neagoe I, Nabavi SM. Habtemariam S, et al. Pharmacol Res. 2020 Jul;157:104853. doi: 10.1016/j.phrs.2020.104853. Epub 2020 Apr 30. Pharmacol Res. 2020. PMID: 32360584 Free PMC article. No abstract available.

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References

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[1] Wu F., et al. , A new coronavirus associated with human respiratory disease in China. Nature, 10.1038/s41586-020-2008-3 (2020). - DOI - PMC - PubMed

[2] Wu F., et al. , A new coronavirus associated with human respiratory disease in China. Nature, 10.1038/s41586-020-2008-3 (2020). - DOI - PMC - PubMed

[3] Zhou P., et al. , A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature, 10.1038/s41586-020-2012-7 (2020). - DOI - PMC - PubMed

[4] Zhou P., et al. , A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature, 10.1038/s41586-020-2012-7 (2020). - DOI - PMC - PubMed

[5] Lu R., et al. , Genomic characterisation and epidemiology of 2019 novel coronavirus: Implications for virus origins and receptor binding. Lancet 395, 565–574 (2020). - PMC - PubMed

[6] Lu R., et al. , Genomic characterisation and epidemiology of 2019 novel coronavirus: Implications for virus origins and receptor binding. Lancet 395, 565–574 (2020). - PMC - PubMed

[7] Zhu N. et al. .; China Novel Coronavirus Investigating and Research Team , A novel coronavirus from patients with pneumonia in China, 2019. N. Engl. J. Med. 382, 727–733 (2020). - PMC - PubMed

[8] Zhu N. et al. .; China Novel Coronavirus Investigating and Research Team , A novel coronavirus from patients with pneumonia in China, 2019. N. Engl. J. Med. 382, 727–733 (2020). - PMC - PubMed

[9] Matsuyama S., et al. , Efficient activation of the severe acute respiratory syndrome coronavirus spike protein by the transmembrane protease TMPRSS2. J. Virol. 84, 12658–12664 (2010). - PMC - PubMed

[10] Matsuyama S., et al. , Efficient activation of the severe acute respiratory syndrome coronavirus spike protein by the transmembrane protease TMPRSS2. J. Virol. 84, 12658–12664 (2010). - PMC - PubMed

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Enhanced isolation of SARS-CoV-2 by TMPRSS2-expressing cells

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Enhanced isolation of SARS-CoV-2 by TMPRSS2-expressing cells

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Abstract

Conflict of interest statement

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