title: Successful gene transfer into dendritic cells with cationized gelatin and plasmid DNA complexes via a phagocytosis-dependent mechanism.
abstract: The use of gene-modified dendritic cells (DC) is a powerful tool to enhance antitumor immune responses stimulated by these cells in cancer immunotherapy. Cationized gelatin is preferably incorporated via phagocytosis and is gradually degraded by proteolysis while buffering lysosomal activity. This may be appropriate for gene transfer into phagocytic cells, such as immature DC. In the present study, successful transfection into monocyte-derived immature DC was demonstrated using cationized gelatin and plasmid DNA complexes. A high transfection efficiency, approaching 16%, was obtained upon transfection of the enhanced green fluorescent protein (EGFP) gene as evaluated by flow cytometry. Transgene expression of EGFP and murine interleukin 12 were also detected by RT-PCR. The antigen-presenting capacity of the transfected DC was equal to that of untransfected DC as evaluated by the allogeneic mixed lymphocyte reaction. Cationized gelatin has the potential to be a unique non-viral vector for gene transfer into DC.
authors.affiliation: Department of Surgery, Division of Digestive Surgery, Kyoto Prefectural University of Medicine, 465 Kajii-cho, Kawaramachi, Hirokoji, Kamigyo-ku, Kyoto 602-8566, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Inada
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Fujiwara
authors.forename: Kiyoto
authors.initials: K
authors.lastname: Atsuji
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Takashima
authors.forename: Yasunobu
authors.initials: Y
authors.lastname: Araki
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kubota
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Tabata
authors.forename: Hisakazu
authors.initials: H
authors.lastname: Yamagishi
mesh_terms: D000818:Animals; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology / Q000502:physiology*; D005780:Gelatin / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D018664:Interleukin-12 / Q000235:genetics; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology / Q000502:physiology; D051379:Mice; D010587:Phagocytosis; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000379:methods*
chemical_list: D049452:Green Fluorescent Proteins; D018664:Interleukin-12; D005780:Gelatin