An HSV amplicon-based helper system for helper-dependent adenoviral vectors.
To produce a helper virus-free stock of helper-dependent adenoviral vectors (HDAdVs), we have developed a new helper system in which adenoviral genes for propagation of HDAdVs are delivered into producer cells by a herpes simplex virus-1 (HSV) amplicon-adenovirus hybrid. The hybrid amplicon was constructed to carry the E1 gene (HA-E1) or the entire adenoviral genome except E1 (HA-Ad). E1 expression from the HSV amplicon successfully complemented propagation of an E1-deleted adenoviral vector in a human glioma cell line. HDAdVs were propagated in 293 cells infected with HA-Ad. In addition, HDAdVs were rescued and propagated in a glioma cell line superinfected with both HA-E1 and HA-Ad amplicons, although relatively low titers of HSV amplicon resulted in low propagation efficiency of HDAdVs. Since the HSV amplicon can be easily and completely inactivated by chloroform extraction and/or heat treatment from the HDAdV stock, this helper system might be an alternative method to produce helper virus-free HDAdVs.
Kubo
Shuji
S
Department of Microbiology, Immunology and Molecular Genetics, UCLA School of Medicine, Los Angeles, CA, USA.
Saeki
Yoshinaga
Y
Chiocca
E Antonio
EA
Mitani
Kohnosuke
K
D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D017498:Adenovirus E1 Proteins / Q000235:genetics; D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D017353:Gene Deletion; D005822:Genetic Vectors; D006378:Helper Viruses / Q000235:genetics*; D018259:Herpesvirus 1, Human / Q000235:genetics*; D006801:Humans; D010957:Plasmids / Q000235:genetics; D014407:Tumor Cells, Cultured; D014709:Vero Cells; D014779:Virus Replication
D017498:Adenovirus E1 Proteins