Am. J. Respir. Cell Mol. Biol.,
Volume 22, Number 2, February, 2000 166-175
Efficient Gene Transfer into Human Normal and Cystic Fibrosis
Tracheal Gland Serous Cells with Synthetic Vectors
Jean-Christophe
Allo,
Patrick
Midoux,
Marc
Merten,
Evelyne
Souil,
Joanna
Lipecka,
Catherine
Figarella,
Michel
Monsigny,
Pascale
Briand,
and
Isabelle
Fajac
INSERM U380, ICGM, Université René Descartes (Paris V), Paris; Centre de Biophysique Moléculaire,
CNRS et Université d'Orléans, Orléans; Groupe de Recherche sur les Glandes Exocrines, Faculté de
Médecine, Marseille; Laboratoire de Physiologie Respiratoire, CHU Cochin, AP-HP-Université
Paris V; and INSERM U467, Faculté Necker, Paris, France
Submucosal gland serous cells are believed to play a major role in the physiopathology of cystic fibrosis
(CF) and may represent an important target for CF gene therapy. We have studied the efficiency of reporter gene transfer into immortalized normal (MM-39) and CF (CF-KM4) human airway epithelial gland
serous cells using various synthetic vectors: glycosylated polylysines (glycofectins), polyethylenimine (PEI) (25 and 800 kD), lipofectin, and lipofectAMINE. In both cell lines, a high luciferase activity was
achieved with various glycofectins, with PEI 25 kD, and with lipofectAMINE. After three transfections
applied daily using 
-glycosylated polylysine, 20% of the cells were transfected. At 24 h after CF transmembrane conductance regulator (CFTR) gene transfer into CF-KM4 cells using -glycosylated polylysine, the immunolocalization of CFTR was analyzed by laser scanning confocal microscopy and the
transgenic CFTR was detected by an intense labeling of the plasma membrane. The presence of membrane
lectins, i.e., cell surface receptors binding oligosaccharides, was also examined on MM-39 and CF-KM4 cells by assessing the binding and uptake of fluorescein-labeled neoglycoproteins and fluorescein-labeled
glycoplexes (glycofectins complexed to plasmid DNA). Among all the neoglycoproteins and glycoplexes
tested, those bearing -mannosylated derivatives were most efficiently taken up by both normal and CF
gland serous cells. However, -mannosylated polylysine was quite inefficient for gene transfer, indicating
that the efficiency of gene transfer is determined both by the uptake of the complexes and also by their intracellular trafficking. Moreover, our results show that an efficient in vitro gene transfer was achieved in
human airway gland serous cells with the same synthetic vectors described to efficiently transfect human
airway surface epithelial cells.
