High-Efficiency Capture of Drug Resistant-Influenza Virus by Live Imaging of Sialidase Activity.

Abstract

Influenza A and B viruses possess a neuraminidase protein that shows sialidase activity. Influenza virus-specific neuraminidase inhibitors (NAIs) are commonly used for clinical treatment of influenza. However, some influenza A and B viruses that are resistant to NAIs have emerged in nature. NAI-resistant viruses have been monitored in public hygiene surveys and the mechanism underlying the resistance has been studied. Here, we describe a new assay for selective detection and isolation of an NAI-resistant virus in a speedy and easy manner by live fluorescence imaging of viral sialidase activity, which we previously developed, in order to achieve high-efficiency capture of an NAI-resistant virus. An NAI-resistant virus maintains sialidase activity even at a concentration of NAI that leads to complete deactivation of the virus. Infected cells and focuses (infected cell populations) of an oseltamivir-resistant virus were selectively visualized by live fluorescence sialidase imaging in the presence of oseltamivir, resulting in high-efficiency isolation of the resistant viruses. The use of a combination of other NAIs (zanamivir, peramivir, and laninamivir) in the imaging showed that the oseltamivir-resistant virus isolated in 2008 was sensitive to zanamivir and laninamivir but resistant to peramivir. Fluorescence imaging in the presence of zanamivir also succeeded in selective live-cell visualization of cells that expressed zanamivir-resistant NA. Fluorescence imaging of NAI-resistant sialidase activity will be a powerful method for study of the NAI resistance mechanism, for public monitoring of NAI-resistant viruses, and for development of a new NAI that shows an effect on various NAI-resistant mutations.

• Authors: Yuuki Kurebayashi, Tadanobu Takahashi, Chihiro Tamoto, Keiji Sahara, Tadamune Otsubo, Tatsuya Yokozawa, Nona Shibahara, Hirohisa Wada, Akira Minami, Kiyoshi Ikeda, Takashi Suzuki
• Journal: PloS one (PLoS One) Vol. 11 Issue 5 Pg. e0156400 ( 2016) ISSN: 1932-6203 [Electronic] United States
• PMID: 27232333 (Publication Type: Journal Article)

Chemical References

• Oseltamivir
• Neuraminidase

Topics

• Animals
• COS Cells
• Cell Survival
• Chlorocebus aethiops
• Dogs
• Drug Resistance, Viral
• Influenza A Virus, H1N1 Subtype (drug effects, enzymology)
• Inhibitory Concentration 50
• Madin Darby Canine Kidney Cells
• Neuraminidase (metabolism)
• Optical Imaging
• Oseltamivir (pharmacology)