Culturing the Unculturable: Human Coronavirus HKU1 Infects, Replicates, and Produces Progeny Virions in Human Ciliated Airway Epithelial Cell Cultures

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  • Krzysztof Pyrc
    • Other Affiliation: Microbiology Department, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland
  • Amy C. Sims
    • Other Affiliation: Department of Epidemiology, School of Public Health, University of North Carolina, Chapel Hill, North Carolina
  • Ronald Dijkman
    • Other Affiliation: Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, Amsterdam, Netherlands
  • Maarten Jebbink
    • Other Affiliation: Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, Amsterdam, Netherlands
  • Casey Long
    • Other Affiliation: Department of Epidemiology, School of Public Health, University of North Carolina, Chapel Hill, North Carolina
  • Damon Deming
    • Other Affiliation: Department of Epidemiology, School of Public Health, University of North Carolina, Chapel Hill, North Carolina
  • Eric Donaldson
    • Other Affiliation: Department of Epidemiology, School of Public Health, University of North Carolina, Chapel Hill, North Carolina
  • Astrid Vabret
    • Other Affiliation: Laboratory of Virology, University Hospital of Caen, Avenue Georges Clemenceau, 14033 Caen Cedex, France
  • Ralph Baric
    • Other Affiliation: Department of Epidemiology, School of Public Health, University of North Carolina, Chapel Hill, North Carolina ; Department of Microbiology and Immunology, School of Medicine, University of North Carolina, Chapel Hill, North Carolina
  • Lia van der Hoek
    • Other Affiliation: Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, Amsterdam, Netherlands
  • Raymond Pickles
    • Other Affiliation: Department of Microbiology and Immunology, School of Medicine, University of North Carolina, Chapel Hill, North Carolina ; Cystic Fibrosis/Pulmonary Research and Treatment Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina
Abstract
  • Culturing newly identified human lung pathogens from clinical sample isolates can represent a daunting task, with problems ranging from low levels of pathogens to the presence of growth suppressive factors in the specimens, compounded by the lack of a suitable tissue culture system. However, it is critical to develop suitable in vitro platforms to isolate and characterize the replication kinetics and pathogenesis of recently identified human pathogens. HCoV-HKU1, a human coronavirus identified in a clinical sample from a patient with severe pneumonia, has been a major challenge for successful propagation on all immortalized cells tested to date. To determine if HCoV-HKU1 could replicate in in vitro models of human ciliated airway epithelial cell cultures (HAE) that recapitulate the morphology, biochemistry, and physiology of the human airway epithelium, the apical surfaces of HAE were inoculated with a clinical sample of HCoV-HKU1 (Cean1 strain). High virus yields were found for several days postinoculation and electron micrograph, Northern blot, and immunofluorescence data confirmed that HCoV-HKU1 replicated efficiently within ciliated cells, demonstrating that this cell type is infected by all human coronaviruses identified to date. Antiserum directed against human leukocyte antigen C (HLA-C) failed to attenuate HCoV-HKU1 infection and replication in HAE, suggesting that HLA-C is not required for HCoV-HKU1 infection of the human ciliated airway epithelium. We propose that the HAE model provides a ready platform for molecular studies and characterization of HCoV-HKU1 and in general serves as a robust technology for the recovery, amplification, adaptation, and characterization of novel coronaviruses and other respiratory viruses from clinical material.
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  • Article
Rights statement
  • In Copyright
Journal title
  • Journal of Virology
Journal volume
  • 84
Journal issue
  • 21
Page start
  • 11255
Page end
  • 11263
Language
  • English
ISSN
  • 0022-538X
  • 1098-5514
  • 1070-6321
Publisher
  • American Society for Microbiology

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