title: Vero/BC-F: an efficient packaging cell line stably expressing F protein to generate single round-infectious human parainfluenza virus type 2 vector.
abstract: A stable packaging cell line (Vero/BC-F) constitutively expressing fusion (F) protein of the human parainfluenza virus type 2 (hPIV2) was established for production of the F-defective and single round-infectious hPIV2 vector in a strategy for recombinant vaccine development. The F gene expression has not evoked cytostatic or cytotoxic effects on the Vero/BC-F cells and the F protein was physiologically active to induce syncytial formation with giant polykaryocytes when transfected with a plasmid expressing hPIV2 hemagglutinin-neuraminidase (HN). Transduction of the F-defective replicon RNA into the Vero/BC-F cells led to the release of the infectious particles that packaged the replicon RNA (named as hPIV2ΔF) without detectable mutations, limiting the infectivity to a single round. The maximal titer of the hPIV2ΔF was 6.0 × 10(8) median tissue culture infections dose per ml. The influenza A virus M2 gene was inserted into hPIV2ΔF, and the M2 protein was found to be highly expressed in a human lung cancer cell line after transduction. Furthermore, in vivo airway infection experiments revealed that the hPIV2ΔF was capable of delivering transgenes to hamster tracheal cells. Thus, non-transmissible or single round-infectious hPIV2 vector will be potentially applicable to human gene therapy or recombinant vaccine development.
authors.affiliation: 1] Department of Microbiology and Molecular Genetics, Mie University Graduate School of Medicine, Tsu, Japan [2] Biocomo Inc., Komono, Komono-cho, Mie, Japan.
authors.forename: J
authors.initials: J
authors.lastname: Ohtsuka
authors.affiliation: 1] Department of Microbiology and Molecular Genetics, Mie University Graduate School of Medicine, Tsu, Japan [2] Biocomo Inc., Komono, Komono-cho, Mie, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Fukumura
authors.affiliation: Department of Microbiology and Molecular Genetics, Mie University Graduate School of Medicine, Tsu, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Tsurudome
authors.affiliation: Department of Microbiology and Molecular Genetics, Mie University Graduate School of Medicine, Tsu, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Hara
authors.affiliation: Department of Microbiology and Molecular Genetics, Mie University Graduate School of Medicine, Tsu, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Nishio
authors.affiliation: Department of Microbiology and Molecular Genetics, Mie University Graduate School of Medicine, Tsu, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Kawano
authors.affiliation: Department of Microbiology and Molecular Genetics, Mie University Graduate School of Medicine, Tsu, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Nosaka
mesh_terms: D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D006224:Cricetinae; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D009980:Influenza A virus / Q000235:genetics; D010223:Parainfluenza Virus 2, Human / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics; D014614:Vaccines, Synthetic / Q000235:genetics; D014709:Vero Cells; D014760:Viral Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D014763:Viral Matrix Proteins / Q000235:genetics; D000086082:Viroporin Proteins
chemical_list: D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D014760:Viral Fusion Proteins; D014763:Viral Matrix Proteins; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins