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 in the development of a gene therapy strategy against human immunodeficiency virus type 1 (HIV-1). Commonly used systems based on retrovirus-mediated gene delivery are characterized by low efficiency gene transfer 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 coupled with DNA-polylysine complexes. Protection of these cells against HIV-1 acute infection was evaluated by transducing them with three different inhibitory genes which interfere with HIV-1 replication at separate levels (polymeric Tat activation response element [TAR] decoy, dominant-negative mutant of the gag gene and antisense sequences of the gag gene) and subsequent challenging with HIV-1. The polymeric TAR decoy inhibited HIV-1 replication over 95%. Both the dominant-negative mutant and the antisense sequence of the gag gene were less potent inhibitors than the polymeric-TAR decoy. Combinations of either polymeric-TAR with dominant-negative mutant or antisense of the gag gene synergistically enhanced the inhibitory effects of the single genes. These data suggest that the combination of a highly efficient transduction technique with effective HIV-1 inhibitory genes confers rapid protection against HIV-1 acute infection in vitro.
Lori
F
F
Laboratory of Tumor Cell Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
Lisziewicz
J
J
Smythe
J
J
Cara
A
A
Bunnag
T A
TA
Curiel
D
D
Gallo
R C
RC
D000256:Adenoviridae / Q000235:genetics; D004274:DNA, Recombinant / Q000008:administration & dosage / Q000235:genetics; D018014:Gene Transfer Techniques; D015750:Genes, gag; D015756:Genes, tat; D015316:Genetic Therapy; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D009154:Mutation; D011107:Polylysine; D013997:Time Factors; D014161:Transduction, Genetic; D014779:Virus Replication / Q000235:genetics*
D004274:DNA, Recombinant; D011107:Polylysine