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Uptake of plasmid/glycosylated polymer complexes and gene transfer efficiency in differentiated airway epithelial cells
Author(s) -
Fajac Isabelle,
Thévenot Guiti,
Bédouet Laurent,
Danel Claire,
Riquet Marc,
Merten Marc,
Figarella Catherine,
AvaSantucci Josette Dall',
Monsigny Michel,
Briand Pascale
Publication year - 2003
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.318
Subject(s) - transfection , flow cytometry , serous fluid , fluorescein , microbiology and biotechnology , biology , gene delivery , cellular differentiation , cell , chemistry , gene , biochemistry , physics , quantum mechanics , fluorescence
Background We have studied gene transfer efficiency of glycosylated polylysines and glycosylated polyethylenimines as vectors in immortalized differentiated airway gland serous cells and primary cultures of human airway surface epithelial cells. Methods In both cell types, lactosylated PEI was more efficient for gene transfer than unsubstituted PEI and lactosylated polylysine which requires the presence of endosomolytic agents. However, for all the vectors tested, gene transfer efficiency was lower in differentiated cells as compared with poorly differentiated cells. The presence of membrane lectins, i.e. cell surface sugar‐specific receptors, was evaluated using fluorescein‐conjugated neoglycoproteins and microscopy or flow cytometry. In differentiated airway surface epithelial cells, membrane lectins were not expressed and plasmid DNA/fluorescein‐conjugated glycosylated polymer complexes were not incorporated. This accounted in part for the lack of gene transfer efficiency in these cells. In contrast, in differentiated airway gland serous cells, expression of lectins and their endocytotic properties appeared to be similar to that observed in undifferentiated cells, and plasmid DNA/fluorescein‐conjugated glycosylated polymer complexes were incorporated in similar amounts by cells in both differentiated states. Results Glycosylated PEI appears to be a promising gene delivery system since it is more efficient than the sugar‐free polymer and does not require endosomolytic agents. However, in differentiated airway gland serous cells, a low gene transfer efficiency was observed that could not be attributed to low expression of membrane lectins or low uptake of glycosylated complexes. An impaired intracellular trafficking of glycosylated complexes in differentiated airway gland serous cells is suggested. Copyright © 2002 John Wiley & Sons, Ltd.