Transfection of human blood monocyte-derived macrophages by transferrin-cationic lipid-DNA complexes
Nejat Düzgüneş: 0000-0001-6159-1391
Gene therapy is emerging as a promising therapeutic strategy for the treatment of genetic metabolic diseases, cancer and AIDS. Macrophages are inflammatory, tumoricidal and microbicidal cells. It may be possible to modulate the function and dysfunction of macrophages by the expression of therapeutic genes. Although liposomal vectors have certain advantages over viral vectors for gene delivery, they are not able to transfect non-dividing cells, such as macrophages. We investigated whether promoting receptormediated endocytosis of cationic liposome-DNA complexes by associating them with transferrin, or by mediating endosome destabilization via fusogenic peptides, would result in transfection of human peripheral blood monocyte-derived macrophages. Complexes composed of DOTAP:DOPE liposomes and pCMVLuc encoding luciferase at different lipid/DNA charge ratios were tested. The association of transferrin with the lipid-DNA complexes resulted in an enhancement of transfection activity for the different charge ratios. The association of the fusogenic peptides "GALA" or HA-2 with the complexes also increased the level of gene expression, but not as effectively as transferrin. The highest levels of transfection were observed when a combination of both transferrin and "GALA" were associated with the complexes. Pre-stimulation of macrophages with hGMCSF prior to the addition of the complexes resulted in an enhancement of transfection for all the conditions described above. The presence of serum did not significantly affect transfection. (JNICT, PRAXIS XXI -BD 4056/94).
Pedroso De Lima, M. C.,
Transfection of human blood monocyte-derived macrophages by transferrin-cationic lipid-DNA complexes.
FASEB Journal, 11(9),