Constitutive expression of chimeric neo-Rev response element transcripts suppresses HIV-1 replication in human CD4+ T lymphocytes.
We have previously reported that chimeric neomycin phosphotransferase (neo)-Rev response element (RRE) transcripts suppress the function of the human immunodeficiency virus type 1 (HIV-1) Rev trans-activator protein in HeLa cells. In an extension of these experiments, human CD4+ CEM cells (G418-resistant cell populations and clonal isolates) stably expressing chimeric neo-RRE genes (2, 3, or 6 RRE copies) were generated using retroviral-mediated gene transfer. The transduced CEM clones were infected with the HIV-1 HTLVIIIB isolate and the following three phenotypes were observed: (i) the transduced CEM cells were readily infected with HIV-1 indistinguishable from the control CEM cells; (ii) the appearance of HIV-1 replication markers was significantly delayed; (iii) no signs of HIV-1 replication were detectable although proviral HIV-1 DNA sequences could be detected in these cells. Furthermore, HIV antigen expression was limited in neo-resistant CEM cell populations inoculated with the HIV-1 HTLVIIIB isolate. Only 10% of the CEM-pX17-3xRRE cells and 20% of the CEM-pX17-2xRRE cells displayed HIV-1 antigens 43 days after challenge and had retained CD4 surface expression on 47% and 64% of the cells, respectively. In sharp contrast, 80% of the CEM-pX17 or the CEM-pX17-6xRRE cells expressed HIV-1 antigens but no CD4 antigens were detectable in these cultures. These results clearly indicate that RRE decoys could be developed into an effective somatic gene therapy approach against HIV-1 induced acquired immunodeficiency syndrome (AIDS).
Bevec
D
D
ART Department, Sandoz Research Institute, Vienna, Austria.
Volc-Platzer
B
B
Zimmermann
K
K
Dobrovnik
M
M
Hauber
J
J
Veres
G
G
Böhnlein
E
E
D001483:Base Sequence; D015704:CD4 Antigens / Q000096:biosynthesis; D002460:Cell Line; D004279:DNA, Viral / Q000302:isolation & purification; D015694:Gene Products, rev / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D015316:Genetic Therapy / Q000379:methods*; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D019868:Kanamycin Kinase; D008969:Molecular Sequence Data; D010641:Phenotype; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000096:biosynthesis / Q000235:genetics; D011533:Proviruses / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D013601:T-Lymphocytes / Q000382:microbiology*; D014162:Transfection; D014779:Virus Replication; D054321:rev Gene Products, Human Immunodeficiency Virus
D015704:CD4 Antigens; D004279:DNA, Viral; D015694:Gene Products, rev; D011993:Recombinant Fusion Proteins; D054321:rev Gene Products, Human Immunodeficiency Virus; D017853:Phosphotransferases (Alcohol Group Acceptor); D019868:Kanamycin Kinase