Intracellular inhibition of HIV-1 replication using a dual protein- and RNA-based strategy.
Exporting unspliced human immunodeficiency virus type 1 (HIV-1) RNA from the nucleus to the cytoplasm, through an interaction between the viral regulatory Rev protein and Rev response element (RRE) RNA, is a critical step in the HIV-1 life-cycle. Disruption of either Rev or the RRE will completely inhibit HIV-1 replication. As such, a strategy for somatic gene therapy to treat HIV-1 infection by intracellular expression of an anti-HIV-1 Rev single chain variable fragment (SFv) and a ribozyme which specifically targets the RRE was developed. The anti-Rev D8SFv, which specifically targets the Rev activation domain, may be a key component of combination intracellular immunization, as it has been previously shown to potently inhibit Rev function, thereby inhibiting viral replication. In the present studies, different HIV-1 RRE region-specific hammerhead ribozymes were constructed and their anti-HIV-1 replication effects were assayed in diverse RNA polymerase (pol) II and III promoters and vector systems in cell culture. Utilizing this combination of an SFv and a ribozyme as a dual strategy to block HIV-1 replication, both at the protein and RNA level, data from these studies demonstrated that potent inhibition of HIV-1 replication can be achieved via this approach. Combination gene therapies hold promise, analogous to combination chemotherapeutic regimens, for the in vivo treatment of HIV-1 infections.
Duan
L
L
Dorrance H Hamilton laboratories, Department of Medicine, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Zhu
M
M
Ozaki
I
I
Zhang
H
H
Wei
D L
DL
Pomerantz
R J
RJ
D001483:Base Sequence; D002478:Cells, Cultured; D003131:Combined Modality Therapy; D015694:Gene Products, rev / Q000276:immunology*; D015752:Genes, env; D015316:Genetic Therapy / Q000379:methods*; D015658:HIV Infections / Q000628:therapy*; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D007128:Immunoglobulin Fragments / Q000627:therapeutic use; D008969:Molecular Sequence Data; D016337:RNA, Catalytic / Q000627:therapeutic use; D013601:T-Lymphocytes / Q000821:virology; D014779:Virus Replication; D054321:rev Gene Products, Human Immunodeficiency Virus
D015694:Gene Products, rev; D007128:Immunoglobulin Fragments; D016337:RNA, Catalytic; D054321:rev Gene Products, Human Immunodeficiency Virus