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Journal of Virology, August 2009, p. 7739-7748, Vol. 83, No. 15
0022-538X/09/$08.00+0     doi:10.1128/JVI.00614-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Human Bocavirus Can Be Cultured in Differentiated Human Airway Epithelial Cells{triangledown}

Ronald Dijkman,1 Sylvie M. Koekkoek,2 Richard Molenkamp,2 Oliver Schildgen,3 and Lia van der Hoek1*

Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands,1 Laboratory of Clinical Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands,2 Institute for Virology, University of Bonn Medical Centre, Bonn, Germany3

Received 25 March 2009/ Accepted 18 May 2009

In 2005, a human bocavirus was discovered in children with respiratory tract illnesses. Attempts to culture this virus on conventional cell lines has failed thus far. We investigated whether the virus can replicate on pseudostratified human airway epithelium. This cell culture system mimics the human airway environment and facilitates culturing of various respiratory agents. The cells were inoculated with human bocavirus-positive nasopharyngeal washes from children, and virus replication was monitored by measuring apical release of the virus via real-time PCR. Furthermore, we identified different viral mRNAs in the infected cells. All mRNAs were transcribed from a single promoter but varied due to alternative splicing and alternative polyadenylation, similar to what has been described for bovine parvovirus and minute virus of canines, the other two members of the Bocavirus genus. Thus, transcription of human bocavirus displays strong homology to the transcription of the other bocaviruses. In conclusion, we report here for the first time that human bocavirus can be propagated in an in vitro culture system and present a detailed map of the set of mRNAs that are produced by the virus.

* Corresponding author. Mailing address: Laboratory of Experimental Virology, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, The Netherlands. Phone: 31 20 566 75 10. Fax: 31 20 691 65 31. E-mail: c.m.vanderhoek{at}amc.uva.nl

{triangledown} Published ahead of print on 27 May 2009.

Journal of Virology, August 2009, p. 7739-7748, Vol. 83, No. 15
0022-538X/09/$08.00+0     doi:10.1128/JVI.00614-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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