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J Virol. Isolation and characterization of current human coronavirus strains in primary human epithelia cultures reveals differences in target cell tropism
[Source: Journal of Virology, full text: (LINK). Abstract, edited.]
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Isolation and characterization of current human coronavirus strains in primary human epithelia cultures reveals differences in target cell tropism
Ronald Dijkman 1,2, Maarten F. Jebbink 2, Sylvie M. Koekkoek 3, Martin Deijs 2, Hulda R. J?nsd?ttir 1, Richard Molenkamp 3, Margareta Ieven 4, Herman Goossens 4, Volker Thiel 1,5 and Lia van der Hoek 2,*
Author Affiliations: <SUP>1</SUP>Institute of Immunobiology, Kantonal Hospital St. Gallen, Switzerland <SUP>2</SUP>Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, The Netherlands <SUP>3</SUP>Laboratory of Clinical Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, The Netherlands <SUP>4</SUP>Department of Medical Microbiology, Vaccine and Infectious Disease Institute, University Hospital Antwerp, Belgium <SUP>5</SUP>Vetsuisse Faculty, University of Zurich, Zurich, Switzerland
ABSTRACT
The human airway epithelium (HAE) represents the entry port of many human respiratory viruses, such as human coronaviruses (HCoVs). Nowadays, four HCoVs, HCoV-229E, HCoV-OC43, HCoV-HKU1 and HCoV-NL63, are known to circulating worldwide, which cause upper and lower respiratory tract infections in non- and hospitalized children. Still studies on fundamental aspects of these HCoV infections at the primary entry port, such as cell tropism are seriously hampered by the lack of a universal culture system, or suitable animal models. To expand the knowledge on fundamental virus-host interactions of all four HCoVs at the site of primary infection, we used pseudo-stratified HAE cell cultures to isolate and characterize clinical representative HCoV strains directly from nasopharyngeal material. Ten contemporary isolates were obtained representing HCoV-229E (n = 1), HCoV-NL63 (n = 1), HCoV-HKU1 (n = 4) and HCoV-OC43 (n = 4). From each strain we analyzed the replication kinetics and progeny virus release on HAE cell cultures derived from different donors. Surprisingly, by visualizing HCoV infection using confocal microscopy, we observed that HCoV-229E employs a target cell tropism for non-ciliated cells whereas HCoV-OC43, HCoV-HKU1 and HCoV-NL63 all infect ciliated cells. Collectively, we demonstrate that HAE cell cultures, that morphologically and functionally resemble human airways in vivo, represent a robust universal culture system to isolate and compare all contemporary HCoV strains.
FOOTNOTES
* Correspondence and requests for material should be addressed to Lia van der Hoek, Laboratory of Experimental Virology, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, The Netherlands; Phone number: +31 20 566 75 10; Fax: +31 20 691 65 31; c.m.vanderhoek@amc.uva.nl
Copyright ? 2013, American Society for Microbiology. All Rights Reserved.
-Ronald Dijkman 1,2, Maarten F. Jebbink 2, Sylvie M. Koekkoek 3, Martin Deijs 2, Hulda R. J?nsd?ttir 1, Richard Molenkamp 3, Margareta Ieven 4, Herman Goossens 4, Volker Thiel 1,5 and Lia van der Hoek 2,*
Author Affiliations: <SUP>1</SUP>Institute of Immunobiology, Kantonal Hospital St. Gallen, Switzerland <SUP>2</SUP>Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, The Netherlands <SUP>3</SUP>Laboratory of Clinical Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, The Netherlands <SUP>4</SUP>Department of Medical Microbiology, Vaccine and Infectious Disease Institute, University Hospital Antwerp, Belgium <SUP>5</SUP>Vetsuisse Faculty, University of Zurich, Zurich, Switzerland
ABSTRACT
The human airway epithelium (HAE) represents the entry port of many human respiratory viruses, such as human coronaviruses (HCoVs). Nowadays, four HCoVs, HCoV-229E, HCoV-OC43, HCoV-HKU1 and HCoV-NL63, are known to circulating worldwide, which cause upper and lower respiratory tract infections in non- and hospitalized children. Still studies on fundamental aspects of these HCoV infections at the primary entry port, such as cell tropism are seriously hampered by the lack of a universal culture system, or suitable animal models. To expand the knowledge on fundamental virus-host interactions of all four HCoVs at the site of primary infection, we used pseudo-stratified HAE cell cultures to isolate and characterize clinical representative HCoV strains directly from nasopharyngeal material. Ten contemporary isolates were obtained representing HCoV-229E (n = 1), HCoV-NL63 (n = 1), HCoV-HKU1 (n = 4) and HCoV-OC43 (n = 4). From each strain we analyzed the replication kinetics and progeny virus release on HAE cell cultures derived from different donors. Surprisingly, by visualizing HCoV infection using confocal microscopy, we observed that HCoV-229E employs a target cell tropism for non-ciliated cells whereas HCoV-OC43, HCoV-HKU1 and HCoV-NL63 all infect ciliated cells. Collectively, we demonstrate that HAE cell cultures, that morphologically and functionally resemble human airways in vivo, represent a robust universal culture system to isolate and compare all contemporary HCoV strains.
FOOTNOTES
* Correspondence and requests for material should be addressed to Lia van der Hoek, Laboratory of Experimental Virology, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, The Netherlands; Phone number: +31 20 566 75 10; Fax: +31 20 691 65 31; c.m.vanderhoek@amc.uva.nl
Copyright ? 2013, American Society for Microbiology. All Rights Reserved.
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