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Synergistic Reactivation of Latent HIV Expression by Ingenol-3-Angelate, PEP005, Targeted NF-kB Signaling in Combination with JQ1 Induced p-TEFb Activation.

Jiang G, Mendes EA, Kaiser P, Wong DP, Tang Y, Cai I, Fenton A, Melcher GP, Hildreth JE, Thompson GR, Wong JK, Dandekar S - PLoS Pathog. (2015)

Bottom Line: Therefore, effective strategies for eradication of the HIV reservoirs are urgently needed.Furthermore, PEP005 and the P-TEFb agonist, JQ1, exhibited synergism in reactivation of latent HIV with a combined effect that is 7.5-fold higher than the effect of PEP005 alone.This anti-cancer compound is a potential candidate for advancing HIV eradication strategies.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Microbiology and Immunology, University of California, Davis, Davis, California, United States of America.

ABSTRACT
Although anti-retroviral therapy (ART) is highly effective in suppressing HIV replication, it fails to eradicate the virus from HIV-infected individuals. Stable latent HIV reservoirs are rapidly established early after HIV infection. Therefore, effective strategies for eradication of the HIV reservoirs are urgently needed. We report that ingenol-3-angelate (PEP005), the only active component in a previously FDA approved drug (PICATO) for the topical treatment of precancerous actinic keratosis, can effectively reactivate latent HIV in vitro and ex vivo with relatively low cellular toxicity. Biochemical analysis showed that PEP005 reactivated latent HIV through the induction of the pS643/S676-PKCδ/θ-IκBα/ε-NF-κB signaling pathway. Importantly, PEP005 alone was sufficient to induce expression of fully elongated and processed HIV RNAs in primary CD4+ T cells from HIV infected individuals receiving suppressive ART. Furthermore, PEP005 and the P-TEFb agonist, JQ1, exhibited synergism in reactivation of latent HIV with a combined effect that is 7.5-fold higher than the effect of PEP005 alone. Conversely, PEP005 suppressed HIV infection of primary CD4+ T cells through down-modulation of cell surface expression of HIV co-receptors. This anti-cancer compound is a potential candidate for advancing HIV eradication strategies.

No MeSH data available.


Related in: MedlinePlus

PEP005 does not induce expression of pro-inflammatory cytokines in primary CD4+ T cells from peripheral blood of HIV-negative donors.CD4+ T cells were isolated from healthy donors and treated with 6 or 12 nM of PEP005 for 24 or 72 hours, and the relative expression of TNF-α (A), IFN-γ (B), IL-2 (C), and IL-6 (D) was quantified using RT-qPCR and normalized to GAPDH internal control.
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ppat.1005066.g006: PEP005 does not induce expression of pro-inflammatory cytokines in primary CD4+ T cells from peripheral blood of HIV-negative donors.CD4+ T cells were isolated from healthy donors and treated with 6 or 12 nM of PEP005 for 24 or 72 hours, and the relative expression of TNF-α (A), IFN-γ (B), IL-2 (C), and IL-6 (D) was quantified using RT-qPCR and normalized to GAPDH internal control.

Mentions: We examined the potential side effects of PEP005 on the inflammatory cytokine expression as a possible consequence of increased CD69 production. It is well recognized that acutely or chronically HIV infected individuals seem to express higher levels of inflammatory cytokines including IL-6 and TNF-α in the peripheral blood [45–47]. Therefore, it is important that the agents for disrupting HIV latency do not exacerbate the unresolved chronic immune activation and inflammatory cytokine expression during HIV eradication interventions. To address this question, we examined CD4+ T cells purified from PBMCs of healthy HIV-negative donors ex vivo for pro-inflammatory cytokine expression following stimulation with PEP005 for 24 or 72 hours. The expression levels of TNF-α, IFN-γ, IL-2, and IL-6 cytokines were determined by RT-qPCR (Fig 6A–6D). There was no significant increase in the expression of these cytokines, except for TNF-α that showed a tendency towards an up-regulation. However, the increase was not statistically significant.


Synergistic Reactivation of Latent HIV Expression by Ingenol-3-Angelate, PEP005, Targeted NF-kB Signaling in Combination with JQ1 Induced p-TEFb Activation.

Jiang G, Mendes EA, Kaiser P, Wong DP, Tang Y, Cai I, Fenton A, Melcher GP, Hildreth JE, Thompson GR, Wong JK, Dandekar S - PLoS Pathog. (2015)

PEP005 does not induce expression of pro-inflammatory cytokines in primary CD4+ T cells from peripheral blood of HIV-negative donors.CD4+ T cells were isolated from healthy donors and treated with 6 or 12 nM of PEP005 for 24 or 72 hours, and the relative expression of TNF-α (A), IFN-γ (B), IL-2 (C), and IL-6 (D) was quantified using RT-qPCR and normalized to GAPDH internal control.
© Copyright Policy
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4520526&req=5

ppat.1005066.g006: PEP005 does not induce expression of pro-inflammatory cytokines in primary CD4+ T cells from peripheral blood of HIV-negative donors.CD4+ T cells were isolated from healthy donors and treated with 6 or 12 nM of PEP005 for 24 or 72 hours, and the relative expression of TNF-α (A), IFN-γ (B), IL-2 (C), and IL-6 (D) was quantified using RT-qPCR and normalized to GAPDH internal control.
Mentions: We examined the potential side effects of PEP005 on the inflammatory cytokine expression as a possible consequence of increased CD69 production. It is well recognized that acutely or chronically HIV infected individuals seem to express higher levels of inflammatory cytokines including IL-6 and TNF-α in the peripheral blood [45–47]. Therefore, it is important that the agents for disrupting HIV latency do not exacerbate the unresolved chronic immune activation and inflammatory cytokine expression during HIV eradication interventions. To address this question, we examined CD4+ T cells purified from PBMCs of healthy HIV-negative donors ex vivo for pro-inflammatory cytokine expression following stimulation with PEP005 for 24 or 72 hours. The expression levels of TNF-α, IFN-γ, IL-2, and IL-6 cytokines were determined by RT-qPCR (Fig 6A–6D). There was no significant increase in the expression of these cytokines, except for TNF-α that showed a tendency towards an up-regulation. However, the increase was not statistically significant.

Bottom Line: Therefore, effective strategies for eradication of the HIV reservoirs are urgently needed.Furthermore, PEP005 and the P-TEFb agonist, JQ1, exhibited synergism in reactivation of latent HIV with a combined effect that is 7.5-fold higher than the effect of PEP005 alone.This anti-cancer compound is a potential candidate for advancing HIV eradication strategies.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Microbiology and Immunology, University of California, Davis, Davis, California, United States of America.

ABSTRACT
Although anti-retroviral therapy (ART) is highly effective in suppressing HIV replication, it fails to eradicate the virus from HIV-infected individuals. Stable latent HIV reservoirs are rapidly established early after HIV infection. Therefore, effective strategies for eradication of the HIV reservoirs are urgently needed. We report that ingenol-3-angelate (PEP005), the only active component in a previously FDA approved drug (PICATO) for the topical treatment of precancerous actinic keratosis, can effectively reactivate latent HIV in vitro and ex vivo with relatively low cellular toxicity. Biochemical analysis showed that PEP005 reactivated latent HIV through the induction of the pS643/S676-PKCδ/θ-IκBα/ε-NF-κB signaling pathway. Importantly, PEP005 alone was sufficient to induce expression of fully elongated and processed HIV RNAs in primary CD4+ T cells from HIV infected individuals receiving suppressive ART. Furthermore, PEP005 and the P-TEFb agonist, JQ1, exhibited synergism in reactivation of latent HIV with a combined effect that is 7.5-fold higher than the effect of PEP005 alone. Conversely, PEP005 suppressed HIV infection of primary CD4+ T cells through down-modulation of cell surface expression of HIV co-receptors. This anti-cancer compound is a potential candidate for advancing HIV eradication strategies.

No MeSH data available.


Related in: MedlinePlus