title: Culturing the unculturable: human coronavirus HKU1 infects, replicates, and produces progeny virions in human ciliated airway epithelial cell cultures.
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.
authors: Microbiology Department, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
authors: Krzysztof
authors: K
authors: Pyrc
authors: Amy C
authors: AC
authors: Sims
authors: Ronald
authors: R
authors: Dijkman
authors: Maarten
authors: M
authors: Jebbink
authors: Casey
authors: C
authors: Long
authors: Damon
authors: D
authors: Deming
authors: Eric
authors: E
authors: Donaldson
authors: Astrid
authors: A
authors: Vabret
authors: Ralph
authors: R
authors: Baric
authors: Lia
authors: L
authors: van der Hoek
authors: Raymond
authors: R
authors: Pickles
mesh_terms: D018929:Cell Culture Techniques; D002478:Cells, Cultured; D017934:Coronavirus / Q000254:growth & development / Q000502:physiology*; D004847:Epithelial Cells / Q000166:cytology / Q000821:virology*; D015236:HLA-C Antigens; D006801:Humans; D008954:Models, Biological; D012137:Respiratory System / Q000473:pathology / Q000821:virology*; D014771:Virion; D053586:Virus Internalization; D014779:Virus Replication
chemical_list: D015236:HLA-C Antigens