GENE-TRANSFER BY DNA GLYCOSYLATED POLYLYSINE COMPLEXES INTO HUMAN BLOOD MONOCYTE-DERIVED MACROPHAGES/

Macrophages are putative target cells for expressing an exogenous gene with therapeutical effects. Knowing that macrophages express membrane lectins mediating endocytosis of their ligands, DNA/glycosylated poly lysine complexes were used to transfect human blood monocyte-derived m acrophages. Monocytes from human peripheral blood were matured in cult ure for 7 days to differentiate into macrophage-like cells in the pres ence of granulocyte-macrophage colony-stimulating factor (GM-CSF). Adh erent cells, which displayed characteristic macrophage markers, CD 14, CD 11b, HLA-DR, and HLA-ABC antigens and mannose receptor, were trans fected by DNA/glycosylated polylysine complexes in the presence of chl oroquine. The luciferase reporter gene expression was maximal 24 hr af ter transfection with a DNA/mannosylated polylysine complex and by usi ng plasmids in which the promoters (either the long terminal repeat of the human immunodeficiency virus or the human cytomegalovirus) drove the luciferase gene expression. Luciferase gene expression was lower w hen the promoter was the early region of the large T antigen of SV40 v irus. Transfection mediated by DNA/mannosylated polylysine complexes w as much more efficient than with DEAE-dextran or lipofectin. The possi bility of transferring and expressing an exogenous gene into macrophag e-like cells by using a nonimmunogenic synthetic vector as a DNA carri er opens new ways to develop nonviral gene therapy strategies.