Coupling of cell-binding ligands to polyethylenimine for targeted gene delivery.
Recently the high transfection potential of the cationic polymer polyethylenimine (PEI) was described (Boussif O et al. Proc Natl Acad Sci USA 1995; 92: 7297-7301). To combine the promising DNA delivering activity of PEI with the concept of receptor-mediated gene delivery, cell-binding ligands (transferrin or antiCD3 antibody) were incorporated by covalent linkage to PEI. DNA complexes of PEI or ligand-PEI conjugates were tested for transfection of cultured neuroblastoma Neuro 2A cells, melanoma B16 or H225 cells, erythroid leukemic K562 cells and T cell leukemia Jurkat E6.1 cells. Depending on the cell line, incorporation of the cell-binding ligand resulted in an up to 1000-fold increased transfection efficiency. This activity depends on ligand-receptor interaction and was observed also at low PEI cation:DNA anion ratios where ligand-free PEI lacks efficiency. Depending on the cell-binding ligand, specific targeting (CD3 antibody, Jurkat cells) can be achieved. Gene transfer can be augmented by the addition of an endosome-destabilizing influenza peptide, but is not dependent on the presence of additional endosomolytic agents. Application of transferrin-PEI for the production of murine interleukin-2 in B16 cells resulted in exceptionally high secretion rates of 19 micrograms IL-2 protein per 10(6) cells per 24 h.
Kircheis
R
R
Boehringer Ingelheim R&D, Vienna, Austria.
Kichler
A
A
Wallner
G
G
Kursa
M
M
Ogris
M
M
Felzmann
T
T
Buchberger
M
M
Wagner
E
E
D000818:Animals; D000906:Antibodies; D001402:B-Lymphocytes / Q000378:metabolism; D017252:CD3 Complex / Q000276:immunology*; D002460:Cell Line; D018390:Gene Targeting; D018014:Gene Transfer Techniques; D006801:Humans; D007376:Interleukin-2 / Q000378:metabolism; D019169:Jurkat Cells; D008024:Ligands; D051379:Mice; D011094:Polyethyleneimine; D011485:Protein Binding; D011990:Receptors, Transferrin / Q000378:metabolism; D014168:Transferrin / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000906:Antibodies; D017252:CD3 Complex; D007376:Interleukin-2; D008024:Ligands; D011990:Receptors, Transferrin; D014168:Transferrin; D011094:Polyethyleneimine