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Journal of Clinical Microbiology, August 2005, p. 4139-4146, Vol. 43, No. 8
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.8.4139-4146.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Enhanced Expression of an {alpha}2,6-Linked Sialic Acid on MDCK Cells Improves Isolation of Human Influenza Viruses and Evaluation of Their Sensitivity to a Neuraminidase Inhibitor

Shuji Hatakeyama,1,2,3 Yuko Sakai-Tagawa,1,2 Maki Kiso,1,2 Hideo Goto,1,2 Chiharu Kawakami,4 Keiko Mitamura,5 Norio Sugaya,6 Yasuo Suzuki,2,7 and Yoshihiro Kawaoka1,2,8*

Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo,1 Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, Saitama,2 Department of Infectious Diseases, Graduate School of Medicine, University of Tokyo, Tokyo,3 Yokohama City Institute of Health, Kanagawa,4 Department of Pediatrics, Kawasaki Municipal Hospital, Kanagawa,5 Department of Pediatrics, Keiyu Hospital, Kanagawa,6 Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences, and COE Program in the 21st Century, Shizuoka, Japan,7 Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, Wisconsin8

Received 15 November 2004/ Returned for modification 28 December 2004/ Accepted 25 April 2005

The extensive use of neuraminidase (NA) inhibitors to treat influenza virus infections mandates close monitoring for resistant variants. Cultured cells do not provide a reliable means of evaluating the susceptibility of human influenza virus isolates to NA inhibitors. That is, the growth of such viruses in cell lines (e.g., Madin-Darby canine kidney [MDCK] cells) is not inhibited by these drugs, even though their sialidase activity is drug-sensitive. Matrosovich et al. (J. Virol. 77:8418-8425, 2003) showed that an MDCK cell line overexpressing the human ß-galactoside {alpha}2,6-sialyltransferase I (ST6Gal I) gene has the potential to assess the sensitivity of human influenza virus isolates to NA inhibitors, based on studies with a limited number of viruses. Here, we asked whether clinical isolates of influenza virus are universally sensitive to an NA inhibitor (oseltamivir) in an MDCK cell line expressing the ST6Gal I gene. The sensitivity of viruses to oseltamivir correlated with the sensitivity of viral sialidase to the compound, demonstrating the potential utility of this modified cell line for detecting NA inhibitor-resistant viruses. Moreover, in ST6Gal I-overexpressing cells, the growth of human influenza viruses was up to 2 logs higher than in MDCK cells. We conclude that the human ST6Gal I-expressing MDCK cell line is useful not only for evaluating their sensitivity to NA inhibitors, but also for isolation of influenza viruses from clinical samples.

* Corresponding author. Mailing address: Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan. Phone: 81-3-5449-5310. Fax: 81-3-5449-5310. E-mail: kawaoka{at}ims.u-tokyo.ac.jp.

Journal of Clinical Microbiology, August 2005, p. 4139-4146, Vol. 43, No. 8
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.8.4139-4146.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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