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Bromelain Inhibits SARS-CoV-2 Infection in VeroE6 Cells

Satish Sagar, Ashok Kumar Rathinavel, William E. Lutz, Lucas R. Struble, Surender Khurana, Andy T Schnaubelt, Nitish Kumar Mishra, Chittibabu Guda, Mara J. Broadhurst, St. Patrick M. Reid, Kenneth W. Bayles, Gloria E. O. Borgstahl, View ORCID ProfilePrakash Radhakrishnan
doi: https://doi.org/10.1101/2020.09.16.297366
Satish Sagar
1Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, Nebraska, USA 68198
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Ashok Kumar Rathinavel
1Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, Nebraska, USA 68198
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William E. Lutz
1Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, Nebraska, USA 68198
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Lucas R. Struble
1Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, Nebraska, USA 68198
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Surender Khurana
2Division of Viral Products, Center for Biologics Evaluation and Research (CBER), FDA, Silver Spring, MD 20871, USA
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Andy T Schnaubelt
3Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, USA 68198
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Nitish Kumar Mishra
4Department of Genetics, Cell Biology and Anatomy, University of Nebraska Medical Center, Omaha, Nebraska, USA 68198
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Chittibabu Guda
4Department of Genetics, Cell Biology and Anatomy, University of Nebraska Medical Center, Omaha, Nebraska, USA 68198
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Mara J. Broadhurst
3Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, USA 68198
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St. Patrick M. Reid
3Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, USA 68198
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Kenneth W. Bayles
3Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, USA 68198
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Gloria E. O. Borgstahl
1Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, Nebraska, USA 68198
5Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center, Omaha, NE, USA 68198
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Prakash Radhakrishnan
1Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, Nebraska, USA 68198
3Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, USA 68198
4Department of Genetics, Cell Biology and Anatomy, University of Nebraska Medical Center, Omaha, Nebraska, USA 68198
5Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center, Omaha, NE, USA 68198
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  • ORCID record for Prakash Radhakrishnan
  • For correspondence: pradhakr@unmc.edu
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Abstract

Coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). The initial interaction between Transmembrane Serine Protease 2 (TMPRSS2) primed SARS-CoV-2 spike (S) protein and host cell receptor angiotensin-converting enzyme 2 (ACE-2) is a pre-requisite step for this novel coronavirus pathogenesis. Here, we expressed a GFP-tagged SARS-CoV-2 S-Ectodomain in Tni insect cells. That contained sialic acid-enriched N- and O-glycans. Surface resonance plasmon (SPR) and Luminex assay showed that the purified S-Ectodomain binding to human ACE-2 and immunoreactivity with COVID-19 positive samples. We demonstrate that bromelain (isolated from pineapple stem and used as a dietary supplement) treatment diminishes the expression of ACE-2 and TMPRSS2 in VeroE6 cells and dramatically lowers the expression of S-Ectodomain. Importantly, bromelain treatment reduced the interaction between S-Ectodomain and VeroE6 cells. Most importantly, bromelain treatment significantly diminished the SARS-CoV-2 infection in VeroE6 cells. Altogether, our results suggest that bromelain or bromelain rich pineapple stem may be used as an antiviral against COVID-19.

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  • Bromelain inhibits / cleaves the expression of ACE-2 and TMPRSS2

  • Bromelain cleaves / degrades SARS-CoV-2 spike protein

  • Bromelain inhibits S-Ectodomain binding and SARS-CoV-2 infection

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Bromelain Inhibits SARS-CoV-2 Infection in VeroE6 Cells
Satish Sagar, Ashok Kumar Rathinavel, William E. Lutz, Lucas R. Struble, Surender Khurana, Andy T Schnaubelt, Nitish Kumar Mishra, Chittibabu Guda, Mara J. Broadhurst, St. Patrick M. Reid, Kenneth W. Bayles, Gloria E. O. Borgstahl, Prakash Radhakrishnan
bioRxiv 2020.09.16.297366; doi: https://doi.org/10.1101/2020.09.16.297366
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Bromelain Inhibits SARS-CoV-2 Infection in VeroE6 Cells
Satish Sagar, Ashok Kumar Rathinavel, William E. Lutz, Lucas R. Struble, Surender Khurana, Andy T Schnaubelt, Nitish Kumar Mishra, Chittibabu Guda, Mara J. Broadhurst, St. Patrick M. Reid, Kenneth W. Bayles, Gloria E. O. Borgstahl, Prakash Radhakrishnan
bioRxiv 2020.09.16.297366; doi: https://doi.org/10.1101/2020.09.16.297366

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