Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2009 Mar 13;284(11):6782-9.
doi: 10.1074/jbc.M807898200. Epub 2009 Jan 9.

Suberoylanilide hydroxamic acid reactivates HIV from latently infected cells

Xavier Contreras  1 , Marc Schweneker, Ching-Shih Chen, Joseph M McCune, Steven G Deeks, Jeffrey Martin, B Matija Peterlin
Affiliations

Suberoylanilide hydroxamic acid reactivates HIV from latently infected cells

Xavier Contreras et al. J Biol Chem. .

Abstract

Human immunodeficiency virus (HIV) persists in a latent form in infected individuals treated effectively with highly active antiretroviral therapy (HAART). In part, these latent proviruses account for the rebound in viral replication observed after treatment interruption. A major therapeutic challenge is to purge this reservoir. In this study, we demonstrate that suberoylanilide hydroxamic acid (SAHA) reactivates HIV from latency in chronically infected cell lines and primary cells. Indeed, P-TEFb, a critical transcription cofactor for HIV, is released and then recruited to the viral promoter upon stimulation with SAHA. The phosphatidylinositol 3-kinase/Akt pathway is involved in the initiation of these events. Using flow cytometry-based single cell analysis of protein phosphorylation, we demonstrate that SAHA activates this pathway in several subpopulations of T cells, including memory T cells that are the major viral reservoir in peripheral blood. Importantly, SAHA activates HIV replication in peripheral blood mononuclear cells from individuals treated effectively with HAART. Thus SAHA, which is a Food and Drug Administration-approved drug, might be considered to accelerate the decay of the latent reservoir in HAART-treated infected humans.

PubMed Disclaimer

Figures

FIGURE 1.
FIGURE 1.
SAHA activates HIV replication at the level of transcription. JΔK (A) or TZM-Bl cells (B) were stimulated with increasing concentrations of SAHA for 48 h. A, viral replication was measured in the supernatant of JΔK cells using p24 ELISA. B, cells were lysed, and luciferase activity was measured. All results are presented as fold induction compared with the unstimulated control. C, PBMCs from healthy donors were infected in vitro with HIV-1LAI, and resting CD4+ T cells were isolated after 14 days of culture in the absence of PHA. These isolated cells were cultured in the presence or absence of SAHA (500 nm) for 2 days, and viral replication was assessed using p24 ELISA. D and E, JΔK and TZM-Bl cells were preincubated or not with Akt and PI3K inhibitors, untreated or treated with SAHA (1 μm), and viral replication was assessed using p24 ELISA.
FIGURE 2.
FIGURE 2.
PI3K/Akt pathway is stimulated by SAHA in primary memory T cells. Comparing stimulated over unstimulated cells, fold changes in phosphorylation was analyzed in T cells (CD3+), CD4+ T cells (CD3+CD4+), naive T cells (CD3+CD4+CD45RA+CD27+), memory T cells (CD3+CD4+CD45RA–), and monocytes/macrophages (CD4+CD33+) for PBMCs of healthy donors. Phosphorylation of Akt (A) and ERK1/2 MAPK (B) was analyzed. Data are representative of three independent experiments conducted with cells from three different donors.
FIGURE 3.
FIGURE 3.
SAHA releases active P-TEFb. Jurkat T cells were untreated or treated with SAHA. At 30 min and 1, 2, or 6 h, cell lysates were subjected to glycerol gradient centrifugation. Fractions were collected and proteins precipitated before Western blotting with anti-HEXIM1 (bottom panels) and anti-Cdk9 (top panels) antibodies. Fractions 1–3 contain the active SC, whereas fractions 5–7 contain the inactive LC.
FIGURE 4.
FIGURE 4.
SAHA stimulates the recruitment of RNAPII and P-TEFb to the HIV LTR. A, schematic representation of the HIV provirus integrated into the genome of JΔK cells. B, JΔK cells were preincubated or not for 1 h with the PI3K inhibitor, LY294002 (10 μm), and then stimulated or not with SAHA (1 μm). ChIP analyses were performed using antibodies against RNAPII and CycT1. The recruitment of these transcription factors to the promoter and coding regions was followed by PCR amplification using primer pairs 1 and 2, respectively.
FIGURE 5.
FIGURE 5.
SAHA activates HIV replication in PBMCs from patients treated effectively with HAART. PBMCs from HAART-treated patients with undetectable viremia for more than 1 year were stimulated or not with SAHA at 500 nm for 6 h (A) or with PHA at 3μg/ml (B) for 24 h. Cells were then cultured in the presence of additional stimulated PBMCs, and viral replication was assessed by p24 ELISA over a period of 21 days. Results from 15 patients are presented. Patients were numbered according to their response to stimuli. Cells from patients 1–10 presented activation of HIV replication, whereas cells from patients 11–15 did not present any HIV replication upon the different stimuli used in these studies.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Chun, T. W., Finzi, D., Margolick, J., Chadwick, K., Schwartz, D., and Siliciano, R. F. (1995) Nat. Med. 1 1284–1290 - PubMed
    1. Finzi, D., Blankson, J., Siliciano, J. D., Margolick, J. B., Chadwick, K., Pierson, T., Smith, K., Lisziewicz, J., Lori, F., Flexner, C., Quinn, T. C., Chaisson, R. E., Rosenberg, E., Walker, B., Gange, S., Gallant, J., and Siliciano, R. F. (1999) Nat. Med. 5 512–517 - PubMed
    1. Lassen, K., Han, Y., Zhou, Y., Siliciano, J., and Siliciano, R. F. (2004) Trends Mol. Med. 10 525–531 - PubMed
    1. Contreras, X., Lenasi, T., and Peterlin, B. M. (2006) Future Virol. 1 733–745 - PMC - PubMed
    1. Lenasi, T., Contreras, X., and Peterlin, B. M. (2008) Cell Host Microbe 4 123–133 - PMC - PubMed

Publication types

MeSH terms

Substances