Specific binding of baculoviruses displaying gp64 fusion proteins to mammalian cells.
Viral vectors displaying specific ligand binding moieties have raised an increasing interest in the area of targeted gene therapy. In this report, we describe baculovirus vectors displaying either a functional single chain antibody fragment (scFv) specific for the carcinoembryonic antigen (CEA) or the synthetic IgG binding domains (ZZ) derived from protein A of Staphylococcus aureus. In addition, the vectors were engineered to incorporate a reporter gene encoding the enhanced green fluorescent protein (EGFP) under the transcriptional regulation of the cytomegalovirus (CMV) IE promoter. Display of the targeting moieties on the viral surface was achieved through fusion to the N-terminus of gp64, the major envelope protein of the Autographa californica nuclear polyhedrosis virus (AcNPV). Specific binding of the gp64 fusion viruses to mammalian target cells was demonstrated by using monoclonal anti-gp64 antibodies followed by fluorescence and/or confocal microscopy. The anti-CEA scFv displaying baculovirus was shown to bind specifically to CEA expressing cells (PC-3). Similarly, the virus displaying the ZZ domains of protein A was targeted to BHK cells via binding of an appropriate IgG antibody. In all cases, the reporter gene was expressed in the transduced mammalian cells as shown by fluorescence microscopy and flow cytometric analyses.
Ojala
K
K
Department of Biological and Environmental Science, Division of Biotechnology, University of Jyväskylä, FIN-40351 Jyväskylä, Finland.
Mottershead
D G
DG
Suokko
A
A
Oker-Blom
C
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics*; D002272:Carcinoembryonic Antigen / Q000276:immunology; D002460:Cell Line; D006224:Cricetinae; D017930:Genes, Reporter; D015316:Genetic Therapy; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology; D008164:Luminescent Proteins / Q000235:genetics; D008856:Microscopy, Fluorescence; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D018411:Spodoptera / Q000235:genetics; D013205:Staphylococcal Protein A / Q000276:immunology; D014161:Transduction, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured; D014760:Viral Fusion Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen; D007135:Immunoglobulin Variable Region; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D014760:Viral Fusion Proteins; D049452:Green Fluorescent Proteins