Human adenovirus serotypes 4p and 11p are efficiently expressed in cell lines of neural tumour origin.
Most currently used adenovirus vectors are based upon adenovirus serotypes 2 and 5 (Ad2 and Ad5), which have limited efficiencies for gene transfer to human neural cells. Both serotypes bind to the known adenovirus receptor, CAR (coxsackievirus and adenovirus receptor), and have restricted cell tropism. The purpose of this study was to find vector candidates that are superior to Ad5 in infecting human neural tumours. Using flow cytometry, the vector candidates Ad4p, Ad11p and Ad17p were compared to the commonly used adenovirus vector Ad5v for their binding capacity to neural cell lines derived from glioblastoma, medulloblastoma and neuroblastoma cell lines. The production of viral structural proteins and the CAR-binding properties of the different serotypes were also assessed in these cells. Computer-based models of the fibre knobs of Ad4p and Ad17 were created based upon the crystallized fibre knob structure of adenoviruses and analysed for putative receptor-interacting regions that differed from the fibre knob of Ad5. The non CAR-binding vector candidate Ad11p showed clearly the best binding capacity to all of the neural cell lines, binding more than 90% of cells of all of the neural cell lines tested, in contrast to 20% or less for the commonly used vector Ad5v. Ad4p and Ad11p were also internalized and produced viral proteins more successfully than Ad5. Ad4p showed a low binding ability but a very efficient capacity for infection in cell culture. Ad17p virions neither bound or efficiently infected any of the neural cell lines studied.
Skog
Johan
J
Department of Virology, Umeå University, SE-901 85 Umeå, Sweden1.
Mei
Ya-Fang
YF
Department of Virology, Umeå University, SE-901 85 Umeå, Sweden1.
Wadell
Göran
G
Department of Virology, Umeå University, SE-901 85 Umeå, Sweden1.
D000260:Adenoviruses, Human / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D002213:Capsid / Q000378:metabolism; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D006224:Cricetinae; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D005909:Glioblastoma; D006801:Humans; D008527:Medulloblastoma; D008969:Molecular Sequence Data; D009447:Neuroblastoma; D011991:Receptors, Virus / Q000378:metabolism; D016415:Sequence Alignment; D012703:Serotyping; D014407:Tumor Cells, Cultured; D015678:Viral Structural Proteins / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*
C568771:CLMP protein, human; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D011991:Receptors, Virus; D015678:Viral Structural Proteins