Structural analysis of vaccinia virus DIs strain: application as a new replication-deficient viral vector.
DIs is a restrictive host range mutant of vaccinia virus strain DIE that grows well only in chick embryo fibroblast cells but is unable to grow in most mammalian cells. In this study, we identified one major deletion (15.4 kbp) which results in the loss of 19 putative open reading frames in the left end of the genome. We then established a system to express foreign genes by inserting them into the deleted region of DIs. We constructed rDIs to express the bacteriophage T7 polymerase (T7pol) gene and showed the expression in various mammalian cell lines by reporter luciferase gene expression under the T7 promoter. We also expressed the full-length human immunodeficiency virus (HIV)-1 NL432 gag gene. The expressed gag gene product induced high levels of cytotoxic T lymphocytes in immunized mice. These data suggest that DIs is useful as an efficient, transient replication-deficient viral vector.
Ishii
Koji
K
Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan.
Ueda
Yoshiaki
Y
Matsuo
Kazuhiro
K
Matsuura
Yoshiharu
Y
Kitamura
Takashi
T
Kato
Kenzo
K
Izumi
Yasuyuki
Y
Someya
Kenji
K
Ohsu
Takeaki
T
Honda
Mitsuo
M
Miyamura
Tatsuo
T
D016915:AIDS Vaccines / Q000008:administration & dosage / Q000276:immunology; D000818:Animals; D017123:Bacteriophage T7 / Q000201:enzymology; D002478:Cells, Cultured; D002642:Chick Embryo; D012321:DNA-Directed RNA Polymerases; D005347:Fibroblasts; D015683:Gene Products, gag / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors; D015497:HIV-1 / Q000235:genetics; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014613:Vaccines, Attenuated / Q000008:administration & dosage / Q000276:immunology; D014616:Vaccinia virus / Q000737:chemistry* / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D014764:Viral Proteins; D014779:Virus Replication
D016915:AIDS Vaccines; D015683:Gene Products, gag; D014613:Vaccines, Attenuated; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases