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Sulforaphane Inhibits HIV Infection of Macrophages through Nrf2.

Furuya AK, Sharifi HJ, Jellinger RM, Cristofano P, Shi B, de Noronha CM - PLoS Pathog. (2016)

Bottom Line: We further found that, like the type I interferon-induced cellular anti-viral proteins SAMHD1 and MX2, SFN treatment blocks infection after entry, but before formation of 2-LTR circles.Interestingly however, neither SAMHD1 nor MX2 were upregulated.This shows for the first time that Nrf2 action can potently block HIV infection and highlights a novel way to trigger this inhibition.

View Article: PubMed Central - PubMed

Affiliation: Center for Immunology and Microbial Disease, Albany Medical College, Albany, New York, United States of America.

ABSTRACT
Marburg virus, the Kaposi's sarcoma-associated herpesvirus (KSHV) and Dengue virus all activate, and benefit from, expression of the transcription regulator nuclear erythroid 2-related factor 2 (Nrf2). The impact of Nrf2 activation on human immunodeficiency virus (HIV) infection has not been tested. Sulforaphane (SFN), produced in cruciferous vegetables after mechanical damage, mobilizes Nrf2 to potently reprogram cellular gene expression. Here we show for the first time that SFN blocks HIV infection in primary macrophages but not in primary T cells. Similarly SFN blocks infection in PMA-differentiated promonocytic cell lines, but not in other cell lines tested. siRNA-mediated depletion of Nrf2 boosted HIV infectivity in primary macrophages and reduced the anti-viral effects of SFN treatment. This supports a model in which anti-viral activity is mediated through Nrf2 after it is mobilized by SFN. We further found that, like the type I interferon-induced cellular anti-viral proteins SAMHD1 and MX2, SFN treatment blocks infection after entry, but before formation of 2-LTR circles. Interestingly however, neither SAMHD1 nor MX2 were upregulated. This shows for the first time that Nrf2 action can potently block HIV infection and highlights a novel way to trigger this inhibition.

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SFN does not trigger expression of interferon-stimulated anti-viral factors SAMHD1 or MX2.PMA-differentiated THP1 cells were mock-treated or treated with media supplemented with vehicle only (DMSO) or with 10 μM SFN or with 500 U/mL of IFNα. (A), Proteins from whole cell lysates were resolved by SDS-PAGE and identified by western blotting using antibodies with the indicated specificities. (B), Densitometric analysis was performed on the Nrf2, SAMHD1, MX2, NQO1 and GCLM (NQO1 and GCLM are both indicators of Nrf2 function) bands and normalized to the values of the corresponding tubulin bands. The relative normalized intensities of the bands were then graphed.
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ppat.1005581.g008: SFN does not trigger expression of interferon-stimulated anti-viral factors SAMHD1 or MX2.PMA-differentiated THP1 cells were mock-treated or treated with media supplemented with vehicle only (DMSO) or with 10 μM SFN or with 500 U/mL of IFNα. (A), Proteins from whole cell lysates were resolved by SDS-PAGE and identified by western blotting using antibodies with the indicated specificities. (B), Densitometric analysis was performed on the Nrf2, SAMHD1, MX2, NQO1 and GCLM (NQO1 and GCLM are both indicators of Nrf2 function) bands and normalized to the values of the corresponding tubulin bands. The relative normalized intensities of the bands were then graphed.

Mentions: Dengue virus infection, in monocyte-derived dendritic cells, activates the IRF3/7/STAT1 and NF-κB anti-viral and inflammatory pathways, as well as Nrf2-dependent antioxidant genes. Interestingly, silencing of Nrf2 with siRNA increased innate anti-viral responses linked to interferon treatment [34]. Our experiments, with HIV in macrophages, sharply contrast this trend. Specifically, boosting Nrf2 counters infection while Nrf2 depletion supports infection. The restriction that we observed however, after reverse transcription but before 2-LTR circle formation, parallels those imposed by the type I interferon-inducible proteins SAMHD1 and MX2 [53, 54]. Here we asked whether SFN-mediated upregulation of Nrf2 protein levels causes an increase in the expression of those anti-viral proteins. We mock treated PMA-differentiated THP1 cultures or treated them with vehicle (DMSO) or 10 μM SFN, or with 500 U/mL of IFNα. As expected, SFN significantly increased levels of Nrf2 as well as of the Nrf2-regulated genes NQO1 and GCLM (Fig 8). SFN did not significantly change levels of either SAMHD1 or MX2. Conversely, IFNα treatment increased levels of MX2 as expected. SAMHD1 levels, despite some donor-to-donor variation, did not change significantly in response to IFNα, nor did the levels of Nrf2, NQO1 or GCLM. Thus, in our system, we do not see an overlap in the set of proteins that are upregulated by SFN and those that are upregulated by IFNα.


Sulforaphane Inhibits HIV Infection of Macrophages through Nrf2.

Furuya AK, Sharifi HJ, Jellinger RM, Cristofano P, Shi B, de Noronha CM - PLoS Pathog. (2016)

SFN does not trigger expression of interferon-stimulated anti-viral factors SAMHD1 or MX2.PMA-differentiated THP1 cells were mock-treated or treated with media supplemented with vehicle only (DMSO) or with 10 μM SFN or with 500 U/mL of IFNα. (A), Proteins from whole cell lysates were resolved by SDS-PAGE and identified by western blotting using antibodies with the indicated specificities. (B), Densitometric analysis was performed on the Nrf2, SAMHD1, MX2, NQO1 and GCLM (NQO1 and GCLM are both indicators of Nrf2 function) bands and normalized to the values of the corresponding tubulin bands. The relative normalized intensities of the bands were then graphed.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4836681&req=5

ppat.1005581.g008: SFN does not trigger expression of interferon-stimulated anti-viral factors SAMHD1 or MX2.PMA-differentiated THP1 cells were mock-treated or treated with media supplemented with vehicle only (DMSO) or with 10 μM SFN or with 500 U/mL of IFNα. (A), Proteins from whole cell lysates were resolved by SDS-PAGE and identified by western blotting using antibodies with the indicated specificities. (B), Densitometric analysis was performed on the Nrf2, SAMHD1, MX2, NQO1 and GCLM (NQO1 and GCLM are both indicators of Nrf2 function) bands and normalized to the values of the corresponding tubulin bands. The relative normalized intensities of the bands were then graphed.
Mentions: Dengue virus infection, in monocyte-derived dendritic cells, activates the IRF3/7/STAT1 and NF-κB anti-viral and inflammatory pathways, as well as Nrf2-dependent antioxidant genes. Interestingly, silencing of Nrf2 with siRNA increased innate anti-viral responses linked to interferon treatment [34]. Our experiments, with HIV in macrophages, sharply contrast this trend. Specifically, boosting Nrf2 counters infection while Nrf2 depletion supports infection. The restriction that we observed however, after reverse transcription but before 2-LTR circle formation, parallels those imposed by the type I interferon-inducible proteins SAMHD1 and MX2 [53, 54]. Here we asked whether SFN-mediated upregulation of Nrf2 protein levels causes an increase in the expression of those anti-viral proteins. We mock treated PMA-differentiated THP1 cultures or treated them with vehicle (DMSO) or 10 μM SFN, or with 500 U/mL of IFNα. As expected, SFN significantly increased levels of Nrf2 as well as of the Nrf2-regulated genes NQO1 and GCLM (Fig 8). SFN did not significantly change levels of either SAMHD1 or MX2. Conversely, IFNα treatment increased levels of MX2 as expected. SAMHD1 levels, despite some donor-to-donor variation, did not change significantly in response to IFNα, nor did the levels of Nrf2, NQO1 or GCLM. Thus, in our system, we do not see an overlap in the set of proteins that are upregulated by SFN and those that are upregulated by IFNα.

Bottom Line: We further found that, like the type I interferon-induced cellular anti-viral proteins SAMHD1 and MX2, SFN treatment blocks infection after entry, but before formation of 2-LTR circles.Interestingly however, neither SAMHD1 nor MX2 were upregulated.This shows for the first time that Nrf2 action can potently block HIV infection and highlights a novel way to trigger this inhibition.

View Article: PubMed Central - PubMed

Affiliation: Center for Immunology and Microbial Disease, Albany Medical College, Albany, New York, United States of America.

ABSTRACT
Marburg virus, the Kaposi's sarcoma-associated herpesvirus (KSHV) and Dengue virus all activate, and benefit from, expression of the transcription regulator nuclear erythroid 2-related factor 2 (Nrf2). The impact of Nrf2 activation on human immunodeficiency virus (HIV) infection has not been tested. Sulforaphane (SFN), produced in cruciferous vegetables after mechanical damage, mobilizes Nrf2 to potently reprogram cellular gene expression. Here we show for the first time that SFN blocks HIV infection in primary macrophages but not in primary T cells. Similarly SFN blocks infection in PMA-differentiated promonocytic cell lines, but not in other cell lines tested. siRNA-mediated depletion of Nrf2 boosted HIV infectivity in primary macrophages and reduced the anti-viral effects of SFN treatment. This supports a model in which anti-viral activity is mediated through Nrf2 after it is mobilized by SFN. We further found that, like the type I interferon-induced cellular anti-viral proteins SAMHD1 and MX2, SFN treatment blocks infection after entry, but before formation of 2-LTR circles. Interestingly however, neither SAMHD1 nor MX2 were upregulated. This shows for the first time that Nrf2 action can potently block HIV infection and highlights a novel way to trigger this inhibition.

Show MeSH
Related in: MedlinePlus