Selective gene delivery in dendritic cells with mannosylated and histidylated lipopolyplexes.
We report for the first time preparation of mannosylated and histidylated lipopolyplexes (Man-LPD100) with uptake and transfection selectivity for dendritic cells (DCs). Man-LPD100 were prepared by addition of mannosylated and histidylated liposomes (Man-Lip100) on preformed PEGylated histidylated polylysine/DNA polyplexes. Man-Lip100 comprised a cationic [O,O-dioleyl-N-(3N-(N-methylimidazolium iodide)propylene) phosphoramidate)] lipid, a neutral [O,O-dioleyl-N-histamine Phosphoramidate] co-lipid and β-D-mannopyranosyl-N-dodecylhexadecanamide (Man-lipid). At the best, Man-Lip100 containing 11 mol % Man-lipid was obtained. We found that dialysis of liposomes completely abolished cytotoxicity. We showed that the uptake of Man(11)-LPD100 by the murine DC line (DC2.4 cells) was at least 10-fold higher than that of Lac(6)-LPD100. A confocal microscopy study with DC2.4 cells expressing Rab5-EGFP or Rab7-EGFP, revealed that DNA uptake occurred through clathrin-mediated endocytosis. The transfection of DC2.4 cells with Man(11)-LPD100 containing DNA encoding luciferase gene gave luciferase activity two to three times higher (9 × 10(5) RLU/mg protein) than with non-mannosylated LPD100. In contrast to the latter, it was inhibited by 90% in the presence of mannose. Overall, the results indicate that mannosylated and histidylated LPD is a promising system for a selective DNA delivery in DCs.
Perche
Federico
F
Centre de Biophysique Moléculaire CNRS UPR, University of Orléans and Inserm, France.
Gosset
David
D
Mével
Mathieu
M
Miramon
Marie-Laure
ML
Yaouanc
Jean-Jacques
JJ
Pichon
Chantal
C
Benvegnu
Thierry
T
Jaffrès
Paul-Alain
PA
Midoux
Patrick
P
D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D003713:Dendritic Cells / Q000378:metabolism*; D004337:Drug Carriers / Q000737:chemistry* / Q000633:toxicity; D004339:Drug Compounding; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D006639:Histidine / Q000737:chemistry*; D008081:Liposomes; D049409:Luciferases, Firefly / Q000235:genetics / Q000378:metabolism; D008358:Mannose / Q000737:chemistry*; D051379:Mice; D018613:Microscopy, Confocal; D010316:Particle Size; D010957:Plasmids; D014162:Transfection; D020691:rab GTP-Binding Proteins / Q000235:genetics; D020696:rab5 GTP-Binding Proteins / Q000235:genetics; D000090802:rab7 GTP-Binding Proteins
D004337:Drug Carriers; D008081:Liposomes; C485184:enhanced green fluorescent protein; D000090802:rab7 GTP-Binding Proteins; C000716465:rab7 GTP-binding proteins, mouse; D049452:Green Fluorescent Proteins; D006639:Histidine; D004247:DNA; D049409:Luciferases, Firefly; D020691:rab GTP-Binding Proteins; D020696:rab5 GTP-Binding Proteins; D008358:Mannose