RGD Inclusion in the Hexon Monomer Provides Adenovirus Type 5-Based Vectors with a Fiber Knob-Independent Pathway for Infection
| Autor: | Emmanuelle Vigne, Michel Perricaudet, Irene Mahfouz, Anne Brie, Patrice Yeh, Jean-François Dedieu |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
medicine.drug_class
Immunoprecipitation viruses Genetic Vectors Molecular Sequence Data Immunology Biology Monoclonal antibody Microbiology Virus Epitope Adenoviridae law.invention Transduction (genetics) Capsid Antigens CD law Virology medicine Humans Amino Acid Sequence HEK 293 cells Gene Therapy Integrin alphaV Molecular biology Insect Science Recombinant DNA Oligopeptides Plasmids |
| Zdroj: | Journal of Virology. 73:5156-5161 |
| ISSN: | 1098-5514 0022-538X |
| DOI: | 10.1128/jvi.73.6.5156-5161.1999 |
| Popis: | Hypervariable region 5 (HVR5) is a hydrophilic, serotypically nonconserved loop of the hexon monomer which extrudes from the adenovirus (Ad) capsid. We have replaced the HVR5 sequence of Ad5 with that of heterologous peptides and studied their effects on virus viability and peptide accessibility. A poliovirus model epitope was first inserted in a series of nine “isogenic” viruses that differed in their flanking spacers. Whereas virus productivity was not profoundly altered by any of these modifications, immunoprecipitation experiments under nondenaturing conditions demonstrated that epitope recognition by its cognate monoclonal antibody (C3 MAb) was strongly linker dependent and correlated perfectly with the ability of C3 MAb to inhibit transgene delivery and expression. An α v -specific ligand (DCRGDCF) was then inserted in a suitable linker context to investigate whether hexon-modified capsids would enhance the transduction of cells displaying limiting amounts of the virus attachment receptors. Interestingly, although hexon has never been implicated in Ad entry, the modified virus significantly increased the transduction of human vascular smooth muscle cells in vitro. Competition experiments with 293 cells saturated with recombinant knob further indicated that the hexon-modified virus could use an additional, knob-independent pathway for entry. We concluded that genetic engineering of the Ad5 hexon monomer constitutes a novel and feasible approach to equip the virus with additional targeting ligands. |
| Databáze: | OpenAIRE |
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