RRC ID |
45131
|
著者 |
Ohtsuka J, Fukumura M, Tsurudome M, Hara K, Nishio M, Kawano M, Nosaka T.
|
タイトル |
Vero/BC-F: an efficient packaging cell line stably expressing F protein to generate single round-infectious human parainfluenza virus type 2 vector.
|
ジャーナル |
Gene Ther
|
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.
|
巻・号 |
21(8)
|
ページ |
775-84
|
公開日 |
2014-8-1
|
DOI |
10.1038/gt.2014.55
|
PII |
gt201455
|
PMID |
24942630
|
MeSH |
Animals
Cell Line
Cells, Cultured
Chlorocebus aethiops
Cricetinae
Gene Transfer Techniques*
Genetic Vectors*
Humans
Influenza A virus / genetics
Parainfluenza Virus 2, Human / genetics*
Recombinant Proteins / genetics
Vaccines, Synthetic / genetics
Vero Cells
Viral Fusion Proteins / genetics*
Viral Fusion Proteins / metabolism
Viral Matrix Proteins / genetics
|
IF |
4.128
|
引用数 |
1
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
GENETICS & HEREDITY
MEDICINE, RESEARCH & EXPERIMENTAL
|
リソース情報 |
ヒト・動物細胞 |
Vero(RCB0001) |