HSV-1 virions engineered for specific binding to cell surface receptors.
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.
Department of Neurology and Department of Radiology, Massachusetts General Hospital, and Neuroscience Program, Harvard Medical School, Boston, MA 02129, USA.
Paola
P
Grandi
Samuel
S
Wang
Deborah
D
Schuback
Victor
V
Krasnykh
Matthew
M
Spear
David T
DT
Curiel
Roberto
R
Manservigi
Xandra O
XO
Breakefield
D000818:Animals; D001667:Binding, Competitive; D015153:Blotting, Western; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002522:Chlorocebus aethiops; D004591:Electrophoresis, Polyacrylamide Gel; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006023:Glycoproteins / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006378:Helper Viruses / Q000235:genetics; D006497:Heparan Sulfate / Q000737:chemistry / Q000378:metabolism; D018259:Herpesvirus 1, Human / Q000235:genetics*; D006801:Humans; D007763:Lac Operon; D008024:Ligands; D008164:Luminescent Proteins / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013997:Time Factors; D014162:Transfection; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000235:genetics*; D014771:Virion / Q000235:genetics*
D006023:Glycoproteins; D049452:Green Fluorescent Proteins; D006497:Heparan Sulfate; D008024:Ligands; D008164:Luminescent Proteins; D014759:Viral Envelope Proteins; C038535:glycoprotein gC, herpes simplex virus type 1