HSV-1 Virions Engineered for Specific Binding to Cell Surface Receptors
| Autor: | Sam Wang, David T. Curiel, Victor Krasnykh, Deborah E. Schuback, Roberto Manservigi, Xandra O. Breakefield, Paola Grandi, Matthew A Spear |
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| Rok vydání: | 2004 |
| Předmět: |
Time Factors
Virion envelope viruses Heparan sulfate Herpesvirus 1 Human Ligands Virus entry Viral Envelope Proteins Genes Reporter Chlorocebus aethiops Drug Discovery Promoter Regions Genetic Cytopathic effect Gene Transfer Techniques Amplicon Lac Operon Helper virus Molecular Medicine Electrophoresis Polyacrylamide Gel Helper Viruses Protein Binding Blotting Western Genetic Vectors Green Fluorescent Proteins Biology Transfection Binding Competitive Virus Cell Line Gene therapy Viral entry Cell surface receptor Genetics Animals Humans Vero Cells Molecular Biology HSV receptors Glycoproteins Pharmacology Targeting Reporter gene Cell Membrane Virion HSV-1 glycoproteins Genetic Therapy Molecular biology Protein Structure Tertiary Luminescent Proteins Heparan sulfate binding Heparitin Sulfate |
| Zdroj: | Molecular Therapy. 9:419-427 |
| ISSN: | 1525-0016 |
| DOI: | 10.1016/j.ymthe.2003.12.010 |
| Popis: | Expression of specific peptide epitopes on the surface of virions has significant potential for studying viral biology and designing vectors for targeted gene therapy. In this study, an HSV-1 amplicon plasmid expressing a modified glycoprotein C (gC), in which the heparan sulfate binding domain was replaced with a His-tag, was used in generating HSV-1 virions. Western blot analysis demonstrated the presence of modified gC in the purified virions. The amplicon vectors were packaged using a gC-, lacZ+ helper virus to generate a mixture of high-titer helper virus (lacZ+) and amplicon vectors (GFP+), which expressed modified gC in the virion envelope. His-tagged virions bound to 293 6H cells expressing a cell surface pseudo-His-tag receptor four-fold more efficiently than to parental 293 cells and also proved more effective than wild-type virus in binding to both cell types. Binding resulted in productive infection by the modified virions with expression of reporter genes and cytopathic effect comparable to those of wild-type virions. Thus, not only can HSV-1 tropism be manipulated to recognize a non-herpes simplex binding receptor, but it is also possible to increase the infective capacity of the vectors beyond that of the wild-type virus via specific ligand receptor combinations. |
| Databáze: | OpenAIRE |
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