title: Complete replication-competent adenovirus 11p vectors with E1 or E3 insertions show improved heat stability.
abstract: Conventional adenovirus vectors harboring E1 or E3 deletions followed by the insertion of an exogenous gene show considerably reduced virion stability. Here, we report strategies to generate complete replication-competent Ad11p(RCAd11p) vectors that overcome the above disadvantage. A GFP cassette was successfully introduced either upstream of E1A or in the E3A region. The resulting vectors showed high expression levels of the hexon and E1genes and also strongly induced the cytopathic effect in targeted cells. When harboring oversized genomes, the RCAd11pE1 and RCAd11pE3 vectors showed significantly improved heat stability in comparison to Ad11pwt;of the three, RCAd11pE3 was the most tolerant to heat treatment. Electron microscopy showed that RCAd11pE3, RCAd11pE1, Ad11pwt, and Ad11pE1 Delmanifested dominant, moderate, minimum, or no full virus particles after heat treatment at 47°C for 5h. Our results demonstrated that both genome size and the insertion site in the viral genome affect virion stability.
authors: Department of Clinical Microbiology and Virology, Umeå University, SE-901 85 Umeå, Sweden. Electronic address: ya-fang.mei@umu.se.
authors: Ya-Fang
authors: YF
authors: Mei
authors: Department of Clinical Microbiology and Virology, Umeå University, SE-901 85 Umeå, Sweden. Electronic address: haidong.wu@umu.se.
authors: Haidong
authors: H
authors: Wu
authors: Division of Clinical Research Centre, Department of Laboratory Medicine, Karolinska Institute, SE-14186 Stockholm, Sweden. Electronic address: kjell.hultenby@ki.se.
authors: Kjell
authors: K
authors: Hultenby
authors: Department of Clinical Microbiology and Virology, Umeå University, SE-901 85 Umeå, Sweden. Electronic address: jim.silver@umu.se.
authors: Jim
authors: J
authors: Silver
mesh_terms: D000260:Adenoviruses, Human / Q000502:physiology* / Q000648:ultrastructure; D001483:Base Sequence; D002460:Cell Line; D003588:Cytopathogenic Effect, Viral; D015870:Gene Expression; D015967:Gene Expression Regulation, Viral; D023061:Gene Order; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics* / Q000648:ultrastructure; D016679:Genome, Viral; D006358:Hot Temperature; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics*; D016254:Mutagenesis, Insertional; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics*
chemical_list: D017874:Immediate-Early Proteins; D014764:Viral Proteins
keywords: Ad11p vectors; Improved stability; Infection; Kinetics; Morphology