Binding of the SARS-CoV-2 Spike Protein to the Asialoglycoprotein Receptor on Human Primary Hepatocytes and Immortalized Hepatocyte-Like Cells by Confocal Analysis

Daniel P Collins,1 Clifford J Steer2,3 1CMDG, LLC, Saint Paul, MN, USA; 2Department of Medicine, University of Minnesota Medical School, Minneapolis, MN, USA; 3Department of Genetics, Cell Biology and Development, University of Minnesota Medical School, Minneapolis, MN, USACorrespondence: Daniel P C...

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Autores principales:	Collins DP, Steer CJ
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2021
Materias:	asialoglycoprotein receptor e12 mlpc sars-cov-2 at2 human hepatocytes spike proteins Diseases of the digestive system. Gastroenterology RC799-869
Acceso en línea:	https://doaj.org/article/08eaa1536cf84db59e8146ba7d199d2f
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spelling	oai:doaj.org-article:08eaa1536cf84db59e8146ba7d199d2f2021-12-02T14:38:58ZBinding of the SARS-CoV-2 Spike Protein to the Asialoglycoprotein Receptor on Human Primary Hepatocytes and Immortalized Hepatocyte-Like Cells by Confocal Analysis1179-1535https://doaj.org/article/08eaa1536cf84db59e8146ba7d199d2f2021-04-01T00:00:00Zhttps://www.dovepress.com/binding-of-the-sars-cov-2-spike-protein-to-the-asialoglycoprotein-rece-peer-reviewed-article-HMERhttps://doaj.org/toc/1179-1535Daniel P Collins,1 Clifford J Steer2,3 1CMDG, LLC, Saint Paul, MN, USA; 2Department of Medicine, University of Minnesota Medical School, Minneapolis, MN, USA; 3Department of Genetics, Cell Biology and Development, University of Minnesota Medical School, Minneapolis, MN, USACorrespondence: Daniel P Collins Email dc@cmdgllc.comBackground: The SARS-CoV-2 virus may have direct or indirect effects on other human organs beyond the respiratory system and including the liver, via binding of the spike protein. This study investigated the potential direct interactions with the liver by comparing the binding of SARS-CoV-2 spike proteins to human AT2-like cells, primary human hepatocytes and immortalized hepatocyte-like hybrid cells. Receptors with binding specificity for SARS-CoV-2 spike protein on AT2 cells and hepatocytes were identified.Methods: The specific binding of biotinylated spike and spike 1 proteins to undifferentiated human E12 MLPC (E12), E12 differentiated alveolar type 2 (AT2) cells, primary human hepatocytes (PHH) and E12 human hepatocyte-like hybrid cells (HLC) was studied by confocal microscopy. We investigated the expression of ACE-2, binding of biotinylated spike protein, biotinylated spike 1 and inhibition of binding by unlabeled spike protein, two neutralizing antibodies and an antibody directed against the hepatocyte asialoglycoprotein receptor 1 (ASGr1).Results: E12 MLPC did not express ACE-2 and did not bind either of spike or spike 1 proteins. AT2-like cells expressed ACE-2 and bound both spike and spike 1. Both PHH and HLC did not express ACE-2 and did not bind spike 1 protein. However, both PHH and HLC actively bound the spike protein. Biotinylated spike protein binding was inhibited by unlabeled spike but not spike 1 protein on PHH and HLC. Two commercial neutralizing antibodies blocked the binding of the spike to PHH and HLC but only one blocked binding to AT2. An antibody to the hepatocyte ASGr1 blocked the binding of the spike protein to PHH and HLC.Conclusion: The absence of ACE-2 receptors and inhibition of spike binding by an antibody to the ASGr1 on both PHH and HLC suggested that the spike protein interacts with the ASGr1. The differential antibody blocking of spike binding to AT2, PHH and HLC indicated that neutralizing activity of SARS-CoV-2 binding might involve additional mechanisms beyond RBD binding to ACE-2.Keywords: asialoglycoprotein receptor, E12 MLPC, SARS-CoV-2, AT2, human hepatocytes, spike proteinsCollins DPSteer CJDove Medical Pressarticleasialoglycoprotein receptore12 mlpcsars-cov-2at2human hepatocytesspike proteinsDiseases of the digestive system. GastroenterologyRC799-869ENHepatic Medicine: Evidence and Research, Vol Volume 13, Pp 37-44 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	asialoglycoprotein receptor e12 mlpc sars-cov-2 at2 human hepatocytes spike proteins Diseases of the digestive system. Gastroenterology RC799-869
spellingShingle	asialoglycoprotein receptor e12 mlpc sars-cov-2 at2 human hepatocytes spike proteins Diseases of the digestive system. Gastroenterology RC799-869 Collins DP Steer CJ Binding of the SARS-CoV-2 Spike Protein to the Asialoglycoprotein Receptor on Human Primary Hepatocytes and Immortalized Hepatocyte-Like Cells by Confocal Analysis
description	Daniel P Collins,1 Clifford J Steer2,3 1CMDG, LLC, Saint Paul, MN, USA; 2Department of Medicine, University of Minnesota Medical School, Minneapolis, MN, USA; 3Department of Genetics, Cell Biology and Development, University of Minnesota Medical School, Minneapolis, MN, USACorrespondence: Daniel P Collins Email dc@cmdgllc.comBackground: The SARS-CoV-2 virus may have direct or indirect effects on other human organs beyond the respiratory system and including the liver, via binding of the spike protein. This study investigated the potential direct interactions with the liver by comparing the binding of SARS-CoV-2 spike proteins to human AT2-like cells, primary human hepatocytes and immortalized hepatocyte-like hybrid cells. Receptors with binding specificity for SARS-CoV-2 spike protein on AT2 cells and hepatocytes were identified.Methods: The specific binding of biotinylated spike and spike 1 proteins to undifferentiated human E12 MLPC (E12), E12 differentiated alveolar type 2 (AT2) cells, primary human hepatocytes (PHH) and E12 human hepatocyte-like hybrid cells (HLC) was studied by confocal microscopy. We investigated the expression of ACE-2, binding of biotinylated spike protein, biotinylated spike 1 and inhibition of binding by unlabeled spike protein, two neutralizing antibodies and an antibody directed against the hepatocyte asialoglycoprotein receptor 1 (ASGr1).Results: E12 MLPC did not express ACE-2 and did not bind either of spike or spike 1 proteins. AT2-like cells expressed ACE-2 and bound both spike and spike 1. Both PHH and HLC did not express ACE-2 and did not bind spike 1 protein. However, both PHH and HLC actively bound the spike protein. Biotinylated spike protein binding was inhibited by unlabeled spike but not spike 1 protein on PHH and HLC. Two commercial neutralizing antibodies blocked the binding of the spike to PHH and HLC but only one blocked binding to AT2. An antibody to the hepatocyte ASGr1 blocked the binding of the spike protein to PHH and HLC.Conclusion: The absence of ACE-2 receptors and inhibition of spike binding by an antibody to the ASGr1 on both PHH and HLC suggested that the spike protein interacts with the ASGr1. The differential antibody blocking of spike binding to AT2, PHH and HLC indicated that neutralizing activity of SARS-CoV-2 binding might involve additional mechanisms beyond RBD binding to ACE-2.Keywords: asialoglycoprotein receptor, E12 MLPC, SARS-CoV-2, AT2, human hepatocytes, spike proteins
format	article
author	Collins DP Steer CJ
author_facet	Collins DP Steer CJ
author_sort	Collins DP
title	Binding of the SARS-CoV-2 Spike Protein to the Asialoglycoprotein Receptor on Human Primary Hepatocytes and Immortalized Hepatocyte-Like Cells by Confocal Analysis
title_short	Binding of the SARS-CoV-2 Spike Protein to the Asialoglycoprotein Receptor on Human Primary Hepatocytes and Immortalized Hepatocyte-Like Cells by Confocal Analysis
title_full	Binding of the SARS-CoV-2 Spike Protein to the Asialoglycoprotein Receptor on Human Primary Hepatocytes and Immortalized Hepatocyte-Like Cells by Confocal Analysis
title_fullStr	Binding of the SARS-CoV-2 Spike Protein to the Asialoglycoprotein Receptor on Human Primary Hepatocytes and Immortalized Hepatocyte-Like Cells by Confocal Analysis
title_full_unstemmed	Binding of the SARS-CoV-2 Spike Protein to the Asialoglycoprotein Receptor on Human Primary Hepatocytes and Immortalized Hepatocyte-Like Cells by Confocal Analysis
title_sort	binding of the sars-cov-2 spike protein to the asialoglycoprotein receptor on human primary hepatocytes and immortalized hepatocyte-like cells by confocal analysis
publisher	Dove Medical Press
publishDate	2021
url	https://doaj.org/article/08eaa1536cf84db59e8146ba7d199d2f
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Binding of the SARS-CoV-2 Spike Protein to the Asialoglycoprotein Receptor on Human Primary Hepatocytes and Immortalized Hepatocyte-Like Cells by Confocal Analysis

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