Isolation of Hokkaido virus, genus Hantavirus, using a newly established cell line derived from the kidney of the grey red-backed vole (Myodes rufocanus bedfordiae).
Hantaviruses belong to the family Bunyaviridae and are maintained in wild rodents. Although Vero E6 cells, which originate from African green monkey kidney, are used widely in hantavirus research, isolation of hantaviruses from this cell line is difficult. To develop an efficient method of propagation and isolation of hantaviruses we established a novel cell line, MRK101, derived from the kidney of the grey red-backed vole (Myodes rufocanus bedfordiae), the natural host of Hokkaido virus (HOKV). The MRK101 cells showed a significantly higher susceptibility to Puumala virus (PUUV) hosted by Myodes glareolus than Vero E6 cells. Viral nucleocapsid protein in PUUV-infected MRK101 cells was detected earlier than in Vero E6 cells, and the viral titre in the culture fluid of MRK101 cells was higher than that of Vero E6 cells during the early phase of infection. In contrast, MRK101 cells showed no susceptibility to Hantaan virus. HOKV, which has not been isolated to date, was isolated successfully using MRK101 cells. Moreover, the newly isolated HOKV was successfully propagated in MRK101, but not Vero E6, cells. Phylogenic analyses of the S (small), M (medium) and L (large) segment sequences revealed that HOKV is related most closely to PUUV, but is distinct from other hantaviruses. These data suggest that the MRK101 cell line is a useful tool for the isolation and propagation of hantaviruses. Moreover, this is (to our knowledge) the first report of hantavirus isolation in a cell line that originated from the natural host.
Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan.
Takahiro
T
Sanada
Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan.
Takahiro
T
Seto
Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan.
Yuka
Y
Ozaki
Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan.
Ngonda
N
Saasa
Graduate School of Medicine, Hokkaido University, Sapporo, Hokkaido 060-0838, Japan.
Kumiko
K
Yoshimatsu
Graduate School of Medicine, Hokkaido University, Sapporo, Hokkaido 060-0838, Japan.
Jiro
J
Arikawa
Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan.
Kentaro
K
Yoshii
Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan.
Hiroaki
H
Kariwa
D000818:Animals; D003411:Arvicolinae / Q000235:genetics / Q000821:virology*; D001483:Base Sequence; D002460:Cell Line; D002522:Chlorocebus aethiops; D006476:Orthohantavirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D007668:Kidney / Q000378:metabolism / Q000821:virology; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins / Q000235:genetics*; D010802:Phylogeny; D029262:Puumala virus / Q000235:genetics; D014709:Vero Cells / Q000821:virology
D019590:Nucleocapsid Proteins