Successful gene transfer into dendritic cells with cationized gelatin and plasmid DNA complexes via a phagocytosis-dependent mechanism.
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.
Inada
Satoshi
S
Department of Surgery, Division of Digestive Surgery, Kyoto Prefectural University of Medicine, 465 Kajii-cho, Kawaramachi, Hirokoji, Kamigyo-ku, Kyoto 602-8566, Japan.
Fujiwara
Hitoshi
H
Atsuji
Kiyoto
K
Takashima
Kazuhiro
K
Araki
Yasunobu
Y
Kubota
Takeshi
T
Tabata
Yasuhiko
Y
Yamagishi
Hisakazu
H
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*
D049452:Green Fluorescent Proteins; D018664:Interleukin-12; D005780:Gelatin