Lactosylated poly-L-lysine targets a potential lactose receptor in cystic fibrosis and non-cystic fibrosis airway epithelial cells.

Poly-L-lysine with 40% of the epsilon -amino groups substituted with lactosyl residues facilitated the internalization of lactosylated poly-L-lysine/cDNA complexes into cystic fibrosis (CF) and non-CF airway epithelial cells. It was previously shown that lactosylated poly-L-lysine enhanced the transfer of cDNA into the cell nucleus, resulting in transfection. The cell entry of lactosylated poly-L-lysine/cDNA complexes, however, has not been elucidated and we hypothesized that entry of the complex was by receptor-mediated endocytosis. It is shown here that binding of the vector/cDNA complexes to the cell membrane was inhibited by lactose but not N-acetyl glucosamine. Examination by electron microscopy revealed the complexes in clathrin-coated pits. Furthermore, the complexes colocalized with transferrin during cell entry and were shown in early endosomes. These results demonstrated that lactosylated poly-L-lysine/cDNA complexes enter airway epithelial cells via receptor-mediated endocytosis utilizing lactose-binding receptors, which employ the clathrin-coated pit for internalization. Taken together with the fact that nuclear translocation also is enhanced by lactose, these results demonstrate why lactosylated poly-L-lysine is an excellent vector for transfection of airway epithelial cells. Moreover, other carbohydrates covalently linked to poly-L-lysine for targeting other specific cell types, combined with lactosyl residues, can be designed for the development of other molecular conjugates for gene transfer.