Involvement of the leader sequence in sendai virus pathogenesis revealed by recovery of a pathogenic field isolate from cDNA

Yutaka Fujii, Takemasa Sakaguchi, Katsuhiro Kiyotani, Cheng Huang, Noriko Fukuhara, Yoshiko Egi, Tetsuya Yoshida

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

We previously demonstrated that a systematic passage of a pathogenic field isolate of Sendai virus (SeV), the Hamamatsu strain, in embryonated eggs caused attenuation of virulence to mice, and we isolated viral clones of distinct virulence (K. Kiyotani et al. Arch. Virol. 146:893-908, 2001). One of the clones, E15cl2, which was obtained from the virus at the 15th egg passage of E0, the parental Hamamatsu clone for egg passage, had 165-fold-attenuated virulence to mice and possessed only four mutations in the entire 15,384-base genome: in an antigenomic sense, U to A at position 20 (U20A) and U to A at position 24 (U24A) in the leader sequence, the promoter for transcription and replication, and A to G at position 9346 (silent) and A to U at position 12174 (Ser to Cys) in the L gene. To examine the possibility that leader mutations affect virus pathogenesis, we recovered live viruses from cDNA derived from the Hamamatsu strain. A mutant virus possessing either a mutation of U20A or U24A in the leader sequence showed a slightly lower pathogenicity than that of the parental virus, whereas a double mutant virus possessing both of the mutations showed 25-fold-attenuated virulence, accompanying a significantly lower virus replication in the mouse lung. Replications of the leader mutant viruses were also impaired in a primary culture of mouse pulmonary epithelial cells but not in chicken embryo fibroblasts. These findings suggest that leader mutations of SeV affect virus pathogenesis by altering virus replication in a host-dependent manner.

Original languageEnglish (US)
Pages (from-to)8540-8547
Number of pages8
JournalJournal of Virology
Volume76
Issue number17
DOIs
StatePublished - Aug 20 2002
Externally publishedYes

Fingerprint

Sendai virus
pathogenesis
Complementary DNA
Viruses
viruses
Virulence
mutation
virulence
Mutation
Clone Cells
mice
clones
Virus Replication
virus replication
mutants
Ovum
lungs
Lung
Eggs
fibroblasts

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Involvement of the leader sequence in sendai virus pathogenesis revealed by recovery of a pathogenic field isolate from cDNA. / Fujii, Yutaka; Sakaguchi, Takemasa; Kiyotani, Katsuhiro; Huang, Cheng; Fukuhara, Noriko; Egi, Yoshiko; Yoshida, Tetsuya.

In: Journal of Virology, Vol. 76, No. 17, 20.08.2002, p. 8540-8547.

Research output: Contribution to journalArticle

Fujii, Yutaka ; Sakaguchi, Takemasa ; Kiyotani, Katsuhiro ; Huang, Cheng ; Fukuhara, Noriko ; Egi, Yoshiko ; Yoshida, Tetsuya. / Involvement of the leader sequence in sendai virus pathogenesis revealed by recovery of a pathogenic field isolate from cDNA. In: Journal of Virology. 2002 ; Vol. 76, No. 17. pp. 8540-8547.
@article{937370da45334e02b87c6831242e7973,
title = "Involvement of the leader sequence in sendai virus pathogenesis revealed by recovery of a pathogenic field isolate from cDNA",
abstract = "We previously demonstrated that a systematic passage of a pathogenic field isolate of Sendai virus (SeV), the Hamamatsu strain, in embryonated eggs caused attenuation of virulence to mice, and we isolated viral clones of distinct virulence (K. Kiyotani et al. Arch. Virol. 146:893-908, 2001). One of the clones, E15cl2, which was obtained from the virus at the 15th egg passage of E0, the parental Hamamatsu clone for egg passage, had 165-fold-attenuated virulence to mice and possessed only four mutations in the entire 15,384-base genome: in an antigenomic sense, U to A at position 20 (U20A) and U to A at position 24 (U24A) in the leader sequence, the promoter for transcription and replication, and A to G at position 9346 (silent) and A to U at position 12174 (Ser to Cys) in the L gene. To examine the possibility that leader mutations affect virus pathogenesis, we recovered live viruses from cDNA derived from the Hamamatsu strain. A mutant virus possessing either a mutation of U20A or U24A in the leader sequence showed a slightly lower pathogenicity than that of the parental virus, whereas a double mutant virus possessing both of the mutations showed 25-fold-attenuated virulence, accompanying a significantly lower virus replication in the mouse lung. Replications of the leader mutant viruses were also impaired in a primary culture of mouse pulmonary epithelial cells but not in chicken embryo fibroblasts. These findings suggest that leader mutations of SeV affect virus pathogenesis by altering virus replication in a host-dependent manner.",
author = "Yutaka Fujii and Takemasa Sakaguchi and Katsuhiro Kiyotani and Cheng Huang and Noriko Fukuhara and Yoshiko Egi and Tetsuya Yoshida",
year = "2002",
month = "8",
day = "20",
doi = "10.1128/JVI.76.17.8540-8547.2002",
language = "English (US)",
volume = "76",
pages = "8540--8547",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "17",

}

TY - JOUR

T1 - Involvement of the leader sequence in sendai virus pathogenesis revealed by recovery of a pathogenic field isolate from cDNA

AU - Fujii, Yutaka

AU - Sakaguchi, Takemasa

AU - Kiyotani, Katsuhiro

AU - Huang, Cheng

AU - Fukuhara, Noriko

AU - Egi, Yoshiko

AU - Yoshida, Tetsuya

PY - 2002/8/20

Y1 - 2002/8/20

N2 - We previously demonstrated that a systematic passage of a pathogenic field isolate of Sendai virus (SeV), the Hamamatsu strain, in embryonated eggs caused attenuation of virulence to mice, and we isolated viral clones of distinct virulence (K. Kiyotani et al. Arch. Virol. 146:893-908, 2001). One of the clones, E15cl2, which was obtained from the virus at the 15th egg passage of E0, the parental Hamamatsu clone for egg passage, had 165-fold-attenuated virulence to mice and possessed only four mutations in the entire 15,384-base genome: in an antigenomic sense, U to A at position 20 (U20A) and U to A at position 24 (U24A) in the leader sequence, the promoter for transcription and replication, and A to G at position 9346 (silent) and A to U at position 12174 (Ser to Cys) in the L gene. To examine the possibility that leader mutations affect virus pathogenesis, we recovered live viruses from cDNA derived from the Hamamatsu strain. A mutant virus possessing either a mutation of U20A or U24A in the leader sequence showed a slightly lower pathogenicity than that of the parental virus, whereas a double mutant virus possessing both of the mutations showed 25-fold-attenuated virulence, accompanying a significantly lower virus replication in the mouse lung. Replications of the leader mutant viruses were also impaired in a primary culture of mouse pulmonary epithelial cells but not in chicken embryo fibroblasts. These findings suggest that leader mutations of SeV affect virus pathogenesis by altering virus replication in a host-dependent manner.

AB - We previously demonstrated that a systematic passage of a pathogenic field isolate of Sendai virus (SeV), the Hamamatsu strain, in embryonated eggs caused attenuation of virulence to mice, and we isolated viral clones of distinct virulence (K. Kiyotani et al. Arch. Virol. 146:893-908, 2001). One of the clones, E15cl2, which was obtained from the virus at the 15th egg passage of E0, the parental Hamamatsu clone for egg passage, had 165-fold-attenuated virulence to mice and possessed only four mutations in the entire 15,384-base genome: in an antigenomic sense, U to A at position 20 (U20A) and U to A at position 24 (U24A) in the leader sequence, the promoter for transcription and replication, and A to G at position 9346 (silent) and A to U at position 12174 (Ser to Cys) in the L gene. To examine the possibility that leader mutations affect virus pathogenesis, we recovered live viruses from cDNA derived from the Hamamatsu strain. A mutant virus possessing either a mutation of U20A or U24A in the leader sequence showed a slightly lower pathogenicity than that of the parental virus, whereas a double mutant virus possessing both of the mutations showed 25-fold-attenuated virulence, accompanying a significantly lower virus replication in the mouse lung. Replications of the leader mutant viruses were also impaired in a primary culture of mouse pulmonary epithelial cells but not in chicken embryo fibroblasts. These findings suggest that leader mutations of SeV affect virus pathogenesis by altering virus replication in a host-dependent manner.

UR - http://www.scopus.com/inward/record.url?scp=0036338003&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036338003&partnerID=8YFLogxK

U2 - 10.1128/JVI.76.17.8540-8547.2002

DO - 10.1128/JVI.76.17.8540-8547.2002

M3 - Article

C2 - 12163573

AN - SCOPUS:0036338003

VL - 76

SP - 8540

EP - 8547

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 17

ER -