Inhibition of HIV type 1 replication in chronically infected monocytes and lymphocytes by retrovirus-mediated gene transfer of anti-Rev single-chain variable fragments.
We investigated a strategy for gene therapy, intracellular expression of anti-HIV-1 Rev single-chain variable fragments (SFvs), in promonocytic (U1) and T (ACH-2) cell lines latently infected with HIV-1. The cellular and molecular mechanisms leading to activation of latent integrated HIV-1 provirus in U1 and ACH-2 cells have been well delineated. These cells produce HIV-1 in response to stimulation with certain cytokines. U1 and ACH-2 cells were transduced with a murine retroviral shuttle vector that expresses anti-Rev SFv (pLXSN-D8SFv-Rev) or with a control murine leukemia virus (MLV) vector (pLXSN). Tumor necrosis factor alpha (TFNalpha)-, interleukin 6 (IL-6)-, and phorbol myristate acid (PMA)-induced HIV-1 expression, as determined by reverse transcriptase (RT) assay, was significantly inhibited in cells transduced with pLXSN-D8SFv-Rev, compared with cells transduced with pLXSN. In addition, pLXSN-D8SFv-Rev-transduced cells, when incubated with monokine-enriched supernatants of human peripheral blood monocyte cultures, produced significantly less HIV-1 than did cells transduced with pLXSN. This resistance to cytokine-induced HIV-1 expression was demonstrated in SFv-transduced U1 and ACH-2 cells maintained in G418-free medium for 2 months. These data suggest that feasibility of utilizing various anti-HIV-1 SFvs to block activation of HIV-1 infection in vivo.
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W Z
WZ
Joseph Stokes Jr. Research Institute of The Children's Hospital of Philadelphia, Department of Pediatrics, University of Pennsylvania Medical School, 19104, USA.
Lai
J P
JP
Bouhamdan
M
M
Duan
L
L
Pomerantz
R J
RJ
Starr
S E
SE
D019380:Anti-HIV Agents / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology; D018929:Cell Culture Techniques; D002460:Cell Line; D015870:Gene Expression; D015694:Gene Products, rev / Q000235:genetics / Q000276:immunology*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D015497:HIV-1 / Q000276:immunology* / Q000502:physiology; D006801:Humans; D007135:Immunoglobulin Variable Region; D015850:Interleukin-6 / Q000494:pharmacology; D009052:Leukemia Virus, Murine; D008214:Lymphocytes / Q000821:virology; D008934:Mitogens / Q000494:pharmacology; D009000:Monocytes / Q000821:virology; D010835:Phytohemagglutinins / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014779:Virus Replication; D054321:rev Gene Products, Human Immunodeficiency Virus
D019380:Anti-HIV Agents; D000911:Antibodies, Monoclonal; D015694:Gene Products, rev; D007135:Immunoglobulin Variable Region; D015850:Interleukin-6; D008934:Mitogens; D010835:Phytohemagglutinins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D014409:Tumor Necrosis Factor-alpha; C092667:anti-HIV-1 Rev SFv; D054321:rev Gene Products, Human Immunodeficiency Virus