Receptor-mediated gene delivery to human peripheral blood mononuclear cells using anti-CD3 antibody coupled to...

Gene transfer to primary cells, especially to lymphoid cells, using a nonviral delivery system has been very challenging. In the present studies, we have used a cationic polymer, polyethylenimine (PEI) coupled to an anti-CD3 antibody for achieving receptor-mediated gene delivery to human peripheral blood mononuclear cells (PBMC). Naive, unstimulated PBMC did not express transfected genes, whereas the transgenes were expressed efficiently in PHA activated PBMC. Transiently expressed gene products were detected maximally at 24 and 48 h following transfection. Gene expression was detected until 96 h with a gradual diminution in the signal after 48 h. Receptor-mediated gene delivery was successfully used for freshly isolated, as well as previously frozen lymphocyte samples. The transfections performed using ligands other than anti-CD3 were not as efficient as anti-CD3-PEI. These results suggest that in addition to receptor-mediated endocytosis, signaling subsequent to engagement of the CD3 receptor with anti-CD3-PEI appears to be important for the efficacy of anti-CD3-PEI mediated gene delivery.