Imaging of influenza virus sialidase activity in living cells.
Influenza virus is rich in variation and mutations. It would be very convenient for virus detection and isolation to histochemically detect viral infection regardless of variation and mutations. Here, we established a histochemical imaging assay for influenza virus sialidase activity in living cells by using a new fluorescent sialidase substrate, 2-(benzothiazol-2-yl)-4-bromophenyl 5-acetamido-3,5-dideoxy-α-D-glycero-D-galacto-2-nonulopyranosidonic acid (BTP3-Neu5Ac). The BTP3-Neu5Ac assay histochemically visualized influenza virus-infected cells regardless of viral hosts and subtypes. Influenza virus neuraminidase-expressed cells, viral focus formation, and virus-infected locations in mice lung tissues were easily, rapidly, and sensitively detected by the BTP3-Neu5Ac assay. Histochemical visualization with the BTP3-Neu5Ac assay is extremely useful for detection of influenza viruses without the need for fixation or a specific antibody. This novel assay should greatly improve the efficiency of detection, titration, and isolation of influenza viruses and might contribute to research on viral sialidase.
1] Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka-shi, Shizuoka 4228526, Japan [2].
Yuuki
Y
Kurebayashi
1] Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka-shi, Shizuoka 4228526, Japan [2].
Tadanobu
T
Takahashi
Department of Organic Chemistry, School of Pharmaceutical Sciences, Hiroshima International University, Kure-shi, Hiroshima 7370112, Japan.
Tadamune
T
Otsubo
Department of Organic Chemistry, School of Pharmaceutical Sciences, Hiroshima International University, Kure-shi, Hiroshima 7370112, Japan.
Kiyoshi
K
Ikeda
Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka-shi, Shizuoka 4228526, Japan.
Shunsaku
S
Takahashi
Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka-shi, Shizuoka 4228526, Japan.
Maiko
M
Takano
Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka-shi, Shizuoka 4228526, Japan.
Takashi
T
Agarikuchi
Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka-shi, Shizuoka 4228526, Japan.
Tsubasa
T
Sato
Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka-shi, Shizuoka 4228526, Japan.
Yukino
Y
Matsuda
Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka-shi, Shizuoka 4228526, Japan.
Akira
A
Minami
Department of Functional Anatomy, School of Nursing, University of Shizuoka, Shizuoka-shi, Shizuoka 4228526, Japan.
Hiroaki
H
Kanazawa
Viral Disease and Epidemiology Research Division, National Institute of Animal Health, National Agriculture and Food Research Organization (NARO), Tsukuba, Ibaraki 3050856, Japan.
Yuko
Y
Uchida
Viral Disease and Epidemiology Research Division, National Institute of Animal Health, National Agriculture and Food Research Organization (NARO), Tsukuba, Ibaraki 3050856, Japan.
Takehiko
T
Saito
Influenza Research Institute, Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, Wisconsin 53711, United States of America; Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo 1088639, Japan; ERATO Infection-Induced Host Responses Project, Japan Science and Technology Agency, Saitama 3320012, Japan.
Yoshihiro
Y
Kawaoka
Shizuoka Institute of Environment and Hygiene, Shizuoka-shi, Shizuoka 4208637, Japan.
Toshihiro
T
Yamada
Shizuoka Institute of Environment and Hygiene, Shizuoka-shi, Shizuoka 4208637, Japan.
Fumihiko
F
Kawamori
Institute for Glycomics, Griffith University, Gold Coast Campus, Queensland 4222, Australia.
Robin
R
Thomson
Institute for Glycomics, Griffith University, Gold Coast Campus, Queensland 4222, Australia.
Mark
M
von Itzstein
Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka-shi, Shizuoka 4228526, Japan.
Takashi
T
Suzuki
D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D004789:Enzyme Activation; D005260:Female; D015870:Gene Expression; D007150:Immunohistochemistry; D051379:Mice; D009439:Neuraminidase / Q000235:genetics / Q000378:metabolism*; D061848:Optical Imaging / Q000379:methods*; D009975:Orthomyxoviridae / Q000201:enzymology* / Q000235:genetics; D010948:Viral Plaque Assay
D009439:Neuraminidase