A comparison of gene transfer methods in human dendritic cells.
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.
Arthur
J F
JF
Division of Surgical Oncology, University of California at Los Angeles School of Medicine 90095-1782, USA.
Butterfield
L H
LH
Roth
M D
MD
Bui
L A
LA
Kiertscher
S M
SM
Lau
R
R
Dubinett
S
S
Glaspy
J
J
McBride
W H
WH
Economou
J S
JS
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
D007376:Interleukin-2; D015851:Interleukin-7