Limits...
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

Expression of T cell activation markers in PEP005-treated primary CD4+ T cells.(A) CD4+ T cells were isolated from uninfected control subjects and treated with 12 nM PEP005 for 24hrs. Total RNA was extracted, and gene expression of CD38, CD69, CD25, or HLA-DR was analyzed by RT-qPCR. (B-F) PBMCs isolated from peripheral blood of healthy HIV-negative donors were treated for 24 or 72 hours with 12 nM of PEP005, and the expression of CD38 (B), CD69 (C, D), and HLA-DR (E, F) was evaluated using flow cytometry after co-staining with the CD3 T cell marker.
© Copyright Policy
Related In: Results  -  Collection

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

ppat.1005066.g005: Expression of T cell activation markers in PEP005-treated primary CD4+ T cells.(A) CD4+ T cells were isolated from uninfected control subjects and treated with 12 nM PEP005 for 24hrs. Total RNA was extracted, and gene expression of CD38, CD69, CD25, or HLA-DR was analyzed by RT-qPCR. (B-F) PBMCs isolated from peripheral blood of healthy HIV-negative donors were treated for 24 or 72 hours with 12 nM of PEP005, and the expression of CD38 (B), CD69 (C, D), and HLA-DR (E, F) was evaluated using flow cytometry after co-staining with the CD3 T cell marker.

Mentions: To be clinically applicable, effective LRAs should be highly potent, minimally cytotoxic and able to penetrate anatomic sanctuaries and immune cell types without inducing global T cell activation [22]. Therefore, we sought to examine the effects of PEP005 on T cell activation and cytotoxicity. Evaluation of the expression of T cell activation biomarkers by RT-qPCR showed that PEP005 treatment did not cause any significant change in the expression of CD38, CD25, or HLA-DR in purified primary CD4+ T cells (Fig 5A). However, there was an increased expression of CD69 in CD4+ T cells. Flow cytometric analysis of global T cells for the expression of CD38, CD69, or HLA-DR further supported the gene expression data. There was no significant change in the expression of CD38 (24 hr) and HLA-DR in CD4+ T cells (Fig 5B, 5E and 5F). However, there was an increase in expression of CD69, an inducible glycoprotein that is expressed early during T lymphocytes activation (Fig 5C and 5D). These findings provide additional evidence that PEP005 reverses HIV latency through PKC-NF-κB signaling in vivo. Since the expression of CD69 is dependent on NF-κB binding to its promoter region, it is understandable that PEP005 induced PKC-NF-κB signaling would up-regulate CD69 expression in CD4+ T cells [44].


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)

Expression of T cell activation markers in PEP005-treated primary CD4+ T cells.(A) CD4+ T cells were isolated from uninfected control subjects and treated with 12 nM PEP005 for 24hrs. Total RNA was extracted, and gene expression of CD38, CD69, CD25, or HLA-DR was analyzed by RT-qPCR. (B-F) PBMCs isolated from peripheral blood of healthy HIV-negative donors were treated for 24 or 72 hours with 12 nM of PEP005, and the expression of CD38 (B), CD69 (C, D), and HLA-DR (E, F) was evaluated using flow cytometry after co-staining with the CD3 T cell marker.
© Copyright Policy
Related In: Results  -  Collection

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

ppat.1005066.g005: Expression of T cell activation markers in PEP005-treated primary CD4+ T cells.(A) CD4+ T cells were isolated from uninfected control subjects and treated with 12 nM PEP005 for 24hrs. Total RNA was extracted, and gene expression of CD38, CD69, CD25, or HLA-DR was analyzed by RT-qPCR. (B-F) PBMCs isolated from peripheral blood of healthy HIV-negative donors were treated for 24 or 72 hours with 12 nM of PEP005, and the expression of CD38 (B), CD69 (C, D), and HLA-DR (E, F) was evaluated using flow cytometry after co-staining with the CD3 T cell marker.
Mentions: To be clinically applicable, effective LRAs should be highly potent, minimally cytotoxic and able to penetrate anatomic sanctuaries and immune cell types without inducing global T cell activation [22]. Therefore, we sought to examine the effects of PEP005 on T cell activation and cytotoxicity. Evaluation of the expression of T cell activation biomarkers by RT-qPCR showed that PEP005 treatment did not cause any significant change in the expression of CD38, CD25, or HLA-DR in purified primary CD4+ T cells (Fig 5A). However, there was an increased expression of CD69 in CD4+ T cells. Flow cytometric analysis of global T cells for the expression of CD38, CD69, or HLA-DR further supported the gene expression data. There was no significant change in the expression of CD38 (24 hr) and HLA-DR in CD4+ T cells (Fig 5B, 5E and 5F). However, there was an increase in expression of CD69, an inducible glycoprotein that is expressed early during T lymphocytes activation (Fig 5C and 5D). These findings provide additional evidence that PEP005 reverses HIV latency through PKC-NF-κB signaling in vivo. Since the expression of CD69 is dependent on NF-κB binding to its promoter region, it is understandable that PEP005 induced PKC-NF-κB signaling would up-regulate CD69 expression in CD4+ T cells [44].

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