RECEPTOR-MEDIATED GENE-TRANSFER INTO HUMAN T-LYMPHOCYTES VIA BINDING OF DNA CD3 ANTIBODY PARTICLES TO THE CD3 T-CELL RECEPTOR COMPLEX/

Retrovirus-mediated gene transfer is currently the method of choice fo r the transfection of human T lymphocytes for applications in gene the rapy. Use of retroviral vectors, however, is hampered by limits on the size of the genetic material to be transferred, the requirement of di viding target cells, and by potential safety questions. Synthetic pept ide-enhanced or adenovirus-enhanced receptor-mediated transferrinfecti on of DNA (SPET and AVET, respectively) is a powerful method for the i ntroduction of genetic material into mammalian cells. Although transfe rrin has proven to be a useful ligand for gene transfer in many cell t ypes, gene expression in T cells with transferrin/DNA complexes is usu ally not satisfactory. To improve gene transfer to T cells, antibodies directed against the CD3-T cell receptor complex were tested for thei r ability to function as ligands for DNA delivery. In T cell lines, up to 50% of the cells expressed a beta-galactosidase reporter gene usin g anti-CD3 gene transfer complexes, Applying optimized conditions, pre stimulated primary peripheral blood lymphocytes were also transfected successfully, although at a lesser efficiency (5%).