Inhibition of human immunodeficiency virus reactivation from latency by a tat transdominant negative mutant.
A BK virus (BKV) expression vector, specific for human cells, was engineered to express antisense human immunodeficiency virus type 1 (HIV-1) tat cDNA (tat-AS) or a tat mutant in cysteine 22 (tat22). Cysteine residues in the cysteine-rich domain of tat are necessary for tat transactivation of the HIV-1 long terminal repeat (LTR). Both the AS tat and the tat mutant significantly inhibited transactivation by tat when assayed in cells cotransfected with an expression vector where the reporter gene for chloramphenicol acetyl transferase was driven by the HIV-1 LTR. Infection of Jurkat cell clones stably expressing tat22 (Jurkat/tat22) or tat-AS (Jurkat/tat-AS) with HIV-1 did not show differences in virus titer in comparison to HIV-1-infected control cells. However, in two Jurkat/tat22 cell clones, entrance of HIV-1 into latency was accelerated significantly and reactivation of HIV-1 from latency induced by tumor necrosis factor-alpha (TNF-alpha) or tat was blocked. These results suggest that, in a combined and integrated approach to the treatment of acquired immunodeficiency syndrome (AIDS), anti-tat genetic therapy could be successfully applied to maintain virus in latency, thereby extending the duration of the asymptomatic phase preceding full-blown AIDS.
Balboni
P G
PG
Institute of Microbiology, School of Medicine, University of Ferrara, Italy.
Bozzini
R
R
Zucchini
S
S
Marconi
P C
PC
Grossi
M P
MP
Caputo
A
A
Manservigi
R
R
Barbanti-Brodano
G
G
D001739:BK Virus; D015152:Blotting, Northern; D015704:CD4 Antigens / Q000032:analysis; D002461:Cell Line, Transformed; D015696:Gene Products, tat / Q000235:genetics; D015756:Genes, tat / Q000235:genetics*; D005822:Genetic Vectors; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology; D006801:Humans; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics; D016372:RNA, Antisense / Q000235:genetics; D012367:RNA, Viral / Q000032:analysis; D014158:Transcription, Genetic / Q000235:genetics; D014775:Virus Activation; D017735:Virus Latency; D054322:tat Gene Products, Human Immunodeficiency Virus
D015704:CD4 Antigens; D015696:Gene Products, tat; D016372:RNA, Antisense; D012367:RNA, Viral; D054322:tat Gene Products, Human Immunodeficiency Virus