An autoregulated dual-function antitat gene for human immunodeficiency virus type 1 gene therapy.
One approach to gene therapy for AIDS is to block the replication of human immunodeficiency virus type 1 (HIV-1) by inhibiting that tat gene, whose product activates the expression of all HIV-1 genes. To accomplish this, we constructed an antitat gene expressing an RNA with dual (polymeric TAR and antisense-tat) function in an attempt to both sequester Tat protein and block its translation from mRNA. A minigene consisting of the antitat gene driven by the HIV-1 long terminal repeat was inserted into a double-copy retrovirus vector, such that antitat expression would be upregulated only in HIV-1-infected cells. After transduction of a T-lymphocytic cell line (Molt-3) the antitat gene inhibited HIV-1 replication. This inhibition was inversely correlated with the virus infections dose. Virus replication was also inhibited for 5 months in two different T-cell lines after they had been infected at a high multiplicity of infection, suggesting that the antitat gene may be effective over long periods. Importantly, antitat blocked the replication and the cytopathic effect of HIV-1 in human peripheral blood mononuclear cells and led to as much as 4,000-fold inhibition of the replication of an HIV-1 field isolate as well as HIV-1 prototypes maintained in culture. These results suggest that antitat gene therapy has potential use for blocking HIV-1 replication in infected individuals.
Lisziewicz
J
J
Laboratory of Tumor Cell Biology, National Cancer Institute, Bethesda, Maryland 20892.
Sun
D
D
Trapnell
B
B
Thomson
M
M
Chang
H K
HK
Ensoli
B
B
Peng
B
B
D000163:Acquired Immunodeficiency Syndrome / Q000628:therapy*; D001483:Base Sequence; D002478:Cells, Cultured; D015967:Gene Expression Regulation, Viral; D018014:Gene Transfer Techniques; D015756:Genes, tat; D015316:Genetic Therapy; D005822:Genetic Vectors; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D013601:T-Lymphocytes / Q000821:virology; D014779:Virus Replication
D009838:Oligodeoxyribonucleotides