title: A tetracycline-regulated cell line produces high-titer lentiviral vectors that specifically target dendritic cells.
abstract: Lentiviral vectors (LVs) are a powerful means of delivering genetic material to many types of cells. Because of safety concerns associated with these HIV-1 derived vectors, producing large quantities of LVs is challenging. In this paper, we report a method for producing high titers of self-inactivating LVs. We retrovirally transduce the tet-off stable producer cell line GPR to generate a cell line, GPRS, which can express all the viral components, including a dendritic cell-specific glycoprotein, SVGmu. Then, we use concatemeric DNA transfection to transfect the LV transfer plasmid encoding a reporter gene GFP in combination with a selectable marker. Several of the resulting clones can produce LV at a titer 10-fold greater than what we achieve with transient transfection. Plus, these viruses efficiently transduce dendritic cells in vitro and generate a strong T cell immune response to our reporter antigen. This method may be a good option for producing strong LV-based vaccines for clinical studies of cancer or infectious diseases.
authors.affiliation: Mork Family Department of Chemical Engineering and Materials Science, Viterbi School of Engineering, University of Southern California, Los Angeles, CA, USA.
authors.forename: Paul D
authors.initials: PD
authors.lastname: Bryson
authors.forename: Chupei
authors.initials: C
authors.lastname: Zhang
authors.forename: Chi-Lin
authors.initials: CL
authors.lastname: Lee
authors.forename: Pin
authors.initials: P
authors.lastname: Wang
mesh_terms: D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000821:virology; D002460:Cell Line; D003713:Dendritic Cells / Q000276:immunology / Q000821:virology*; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics* / Q000276:immunology; D010957:Plasmids / Q000235:genetics; D013752:Tetracycline / Q000494:pharmacology*; D014162:Transfection / Q000379:methods*
chemical_list: D013752:Tetracycline