RAPID PROTECTION AGAINST HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 (HIV-1) REPLICATION MEDIATED BY HIGH-EFFICIENCY NON-RETROVIRAL DELIVERY OF GENES INTERFERING WITH HIV-1 TAT AND GAG

Efficient transduction of inhibitory genes is a critical requirement i n the development of a gene therapy strategy against human immunodefic iency virus type 1 (HIV-1). Commonly used systems based on retrovirus- mediated gene delivery are characterized by low efficiency gene transf er into the target cell. Genes were transduced in the absence of cell selection into 60-90% of human CD4(+) cells by using a novel technique that allows high efficiency gene transfer mediated by adenoviruses co upled with DNA-polylysine complexes. Protection of these cells against HIV-1 acute infection was evaluated by transducing them with three di fferent inhibitory genes which interfere with HIV-1 replication at sep arate levels (polymeric Tat activation response element [TAR] decoy, d ominant-negative mutant of the gag gene and antisense sequences of the gag gene) and subsequent challenging with HIV-1 replication over 95%. Both the dominant-negative mutant and the antisense sequence of the g ag gene were less potent inhibitors than the polymeric-TAR decoy. Comb inations of either polymeric-TAR with dominant-negative mutant or anti sense of the gag gene synergistically enhanced the inhibitory effects of the single genes. These data suggest that the combination of a high ly efficient transduction technique with effective HIV-1 inhibitory ge nes confers rapid protection against HIV-1 acute infection in vitro.