title: A comparison of gene transfer methods in human dendritic cells.
abstract: Dendritic cells (DCs) are the most potent antigen-presenting cells (APCs) for the initiation of antigen-specific T-cell activation. DCs may be highly enriched from peripheral blood-adherent leukocytes by short-term (7-day) culture in the presence of interleukin (IL)-4 and granulocyte-macrophage colony-stimulating factor. Various methods of gene transfer were studied, including DNA/liposome complexes, electroporation, CaPO4 precipitation, and recombinant adenovirus (AdV) vectors. Low levels of expression were obtained with the physical methods tested. In contrast, AdV vectors expressing luciferase, beta-galactosidase, IL-2, and IL-7 all readily transduced human DCs. Increasing levels of gene expression were observed over a range of multiplicity of infection (MOI) of 10:1 to 10,000:1, with transduction efficiencies exceeding 95% at higher MOI. Although levels of maximal gene expression in DCs were significantly lower than those obtained using human tumor cell lines, IL-2 and IL-7 production of up to 5 x 10(2) ng/10(6) DC were achieved. These results suggest that AdV vectors are a promising vehicle for genetically engineering human DCs.
authors.affiliation: Division of Surgical Oncology, University of California at Los Angeles School of Medicine 90095-1782, USA.
authors.forename: J F
authors.initials: JF
authors.lastname: Arthur
authors.forename: L H
authors.initials: LH
authors.lastname: Butterfield
authors.forename: M D
authors.initials: MD
authors.lastname: Roth
authors.forename: L A
authors.initials: LA
authors.lastname: Bui
authors.forename: S M
authors.initials: SM
authors.lastname: Kiertscher
authors.forename: R
authors.initials: R
authors.lastname: Lau
authors.forename: S
authors.initials: S
authors.lastname: Dubinett
authors.forename: J
authors.initials: J
authors.lastname: Glaspy
authors.forename: W H
authors.initials: WH
authors.lastname: McBride
authors.forename: J S
authors.initials: JS
authors.lastname: Economou
mesh_terms: D003713:Dendritic Cells / Q000276:immunology; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D006801:Humans; D016130:Immunophenotyping; D007376:Interleukin-2 / Q000096:biosynthesis / Q000235:genetics; D015851:Interleukin-7 / Q000096:biosynthesis / Q000235:genetics; D008722:Methods; D011995:Recombination, Genetic
chemical_list: D007376:Interleukin-2; D015851:Interleukin-7