title: Efficient foreign gene expression in Epstein-Barr virus-transformed human B-cells.
abstract: Epstein-Barr virus (EBV) is a herpesvirus that transforms B-cells (B-LCL) and has undergone intense scrutiny owing to its association with Burkitt's lymphoma, nasopharyngeal carcinoma, and immunoblastic lymphomas. B-LCL have also proven useful in the study of human immunology. We describe a novel system for inducing efficient foreign gene expression in B-LCL using biotinylated adenovirus as an endosome-disrupting agent. Plasmid DNA is coupled to the exterior of viral particles by streptavidin-polylysine chimeric proteins. Up to 67% of B-LCL may be induced to express foreign genes in vitro in transient expression systems, and gene expression lasts for at least 17 days. We have expressed firefly luciferase, beta-galactosidase (beta-gal), chloramphenicol acetyltransferase, HIV gag, and env genes, as well as infectious HIV, and the EBV-specific BZLF gene in B-LCL with this system. In vivo delivery of a beta-gal reporter gene to B-LCL was documented in a SCID mouse model. Potential applications include study of genetic regulation of EBV infection and transformation events, study of potential gene therapies for EBV-related B-cell tumors, and production of antigen-presenting cells for use in immunologic assays. Because of the high percentage of cells transformed and the length of foreign gene expression, the possibility of examining foreign gene expression in transient assays, without selection for clonal populations, exists.
authors.affiliation: Division of Infectious Disease, University of Colorado Health Sciences Center, Denver 80262.
authors.forename: T J
authors.initials: TJ
authors.lastname: Curiel
authors.forename: D R
authors.initials: DR
authors.lastname: Cook
authors.forename: C
authors.initials: C
authors.lastname: Bogedain
authors.forename: W
authors.initials: W
authors.lastname: Jilg
authors.forename: G S
authors.initials: GS
authors.lastname: Harrison
authors.forename: M
authors.initials: M
authors.lastname: Cotten
authors.forename: D T
authors.initials: DT
authors.lastname: Curiel
authors.forename: E
authors.initials: E
authors.lastname: Wagner
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000235:genetics; D001402:B-Lymphocytes / Q000382:microbiology*; D002472:Cell Transformation, Viral / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics; D019309:Epstein-Barr Virus Nuclear Antigens; D005260:Female; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics*; D006678:HIV / Q000235:genetics; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D008297:Male; D051379:Mice; D016513:Mice, SCID; D011401:Promoter Regions, Genetic / Q000235:genetics; D011961:Receptors, Fc / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014162:Transfection / Q000379:methods; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D000956:Antigens, Viral; D004268:DNA-Binding Proteins; D019309:Epstein-Barr Virus Nuclear Antigens; D011961:Receptors, Fc; D011994:Recombinant Proteins; D014764:Viral Proteins