Synergistic activation of HIV-1 expression by deacetylase inhibitors and prostratin: implications for treatment of latent infection.
The persistence of transcriptionally silent but replication-competent HIV-1 reservoirs in Highly Active Anti-Retroviral Therapy (HAART)-treated infected individuals, represents a major hurdle to virus eradication. Activation of HIV-1 gene expression in these cells together with an efficient HAART has been proposed as an adjuvant therapy aimed at decreasing the pool of latent viral reservoirs. Using the latently-infected U1 monocytic cell line and latently-infected J-Lat T-cell clones, we here demonstrated a strong synergistic activation of HIV-1 production by clinically used histone deacetylase inhibitors (HDACIs) combined with prostratin, a non-tumor-promoting nuclear factor (NF)- kappaB inducer. In J-Lat cells, we showed that this synergism was due, at least partially, to the synergistic recruitment of unresponsive cells into the expressing cell population. A combination of prostratin+HDACI synergistically activated the 5' Long Terminal Repeat (5'LTR) from HIV-1 Major group subtypes representing the most prevalent viral genetic forms, as shown by transient transfection reporter assays. Mechanistically, HDACIs increased prostratin-induced DNA-binding activity of nuclear NF-kappaB and degradation of cytoplasmic NF-kappaB inhibitor, IkappaBalpha . Moreover, the combined treatment prostratin+HDACI caused a more pronounced nucleosomal remodeling in the U1 viral promoter region than the treatments with the compounds alone. This more pronounced remodeling correlated with a synergistic reactivation of HIV-1 transcription following the combined treatment prostratin+HDACI, as demonstrated by measuring recruitment of RNA polymerase II to the 5'LTR and both initiated and elongated transcripts. The physiological relevance of the prostratin+HDACI synergism was shown in CD8(+)-depleted peripheral blood mononuclear cells from HAART-treated patients with undetectable viral load. Moreover, this combined treatment reactivated viral replication in resting CD4(+) T cells isolated from similar patients. Our results suggest that combinations of different kinds of proviral activators may have important implications for reducing the size of latent HIV-1 reservoirs in HAART-treated patients.
Reuse
Sophie
S
Laboratory of Molecular Virology, Institut de Biologie et de Médecine Moléculaires (IBMM), Université Libre de Bruxelles (ULB), Gosselies, Belgium.
Calao
Miriam
M
Kabeya
Kabamba
K
Guiguen
Allan
A
Gatot
Jean-Stéphane
JS
Quivy
Vincent
V
Vanhulle
Caroline
C
Lamine
Aurélia
A
Vaira
Dolores
D
Demonte
Dominique
D
Martinelli
Valérie
V
Veithen
Emmanuelle
E
Cherrier
Thomas
T
Avettand
Véronique
V
Poutrel
Solène
S
Piette
Jacques
J
de Launoit
Yvan
Y
Moutschen
Michel
M
Burny
Arsène
A
Rouzioux
Christine
C
De Wit
Stéphane
S
Herbein
Georges
G
Rohr
Olivier
O
Collette
Yves
Y
Lambotte
Olivier
O
Clumeck
Nathan
N
Van Lint
Carine
C
D000328:Adult; D000368:Aged; D019380:Anti-HIV Agents / Q000494:pharmacology*; D018414:CD8-Positive T-Lymphocytes / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004357:Drug Synergism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D015658:HIV Infections / Q000188:drug therapy*; D015497:HIV-1 / Q000201:enzymology* / Q000378:metabolism*; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D008875:Middle Aged; D009000:Monocytes / Q000821:virology; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D009707:Nucleosomes / Q000378:metabolism; D010703:Phorbol Esters / Q000494:pharmacology*; D017735:Virus Latency / Q000187:drug effects*
D019380:Anti-HIV Agents; D004791:Enzyme Inhibitors; D020672:I-kappa B Proteins; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D009707:Nucleosomes; D010703:Phorbol Esters; D000072000:NF-KappaB Inhibitor alpha; C070999:prostratin