Lactosylated polylysine, an effective vector to deliver reporter genes into airway epithelial cells is non immunogenic and therefore has the potential of providing an alternative to viral vectors and liposomes(1). Transfection efficiency was assessed using the reporter gene LacZ and glycerol during transfection. In some cases a high dose of vector/plasmid complex may not be feasible because of concentration and toxicity in vivo. Therefore lower doses given sequentially were examined. To assay gene transfection, lactosylated polylysine was complexed to pCMVLacZ in a 3/1 (w/w) ratio. Transfection was performed with immortalized CF airway epithelial cells (CF/T43) in the presence of 100 μM chloroquine and 1% FBS for 4 h at 37°C. After 48 h expression time, the cells were stained with X-GAL for 18 h and the blue cells were counted. The use of 5% glycerol in the presence of 100 μM chloroquine increased transfection efficiency threefold. A dose of 6 μg plasmid one time or 2 μg plasmid on three sequential days in appropriate complex with lactosylated polylysine, resulted in 80%-90% transfection efficiency. Therefore, lactosylated polylysine is a highly efficient vector for gene transfer into CF airway epithelial cells. These non viral vectors were also successful for gene transfer into non CF airway epithelial cells. The use of glycerol during transfection and repeated administration of the vector-plasmid complex will be of special interest in the transfer of CFTR into airway epithelial cells in vivo.

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