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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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Related in: MedlinePlus

The SFN-mediated HIV infection block is reversible.(A), Study design: PMA-differentiated THP1 cells were pretreated with media supplemented with vehicle only (DMSO) or with 10 μM, 20 μM or 30 μM SFN. After twenty-four hours of treatment, the culture media was replaced with fresh SFN-free media. Cultures were infected 1, 2, 3, 4, or 5 days after treatment with freshly thawed aliquots of VSV-G pseudotyped HIV-1 encoding firefly luciferase in place of nef. Twenty-four hours after the start of each time-point infection, the cells were harvested and (B), luciferase activity was measured by photon emission. The bar graph represents the data for replicate experiments (n = 3).
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ppat.1005581.g003: The SFN-mediated HIV infection block is reversible.(A), Study design: PMA-differentiated THP1 cells were pretreated with media supplemented with vehicle only (DMSO) or with 10 μM, 20 μM or 30 μM SFN. After twenty-four hours of treatment, the culture media was replaced with fresh SFN-free media. Cultures were infected 1, 2, 3, 4, or 5 days after treatment with freshly thawed aliquots of VSV-G pseudotyped HIV-1 encoding firefly luciferase in place of nef. Twenty-four hours after the start of each time-point infection, the cells were harvested and (B), luciferase activity was measured by photon emission. The bar graph represents the data for replicate experiments (n = 3).

Mentions: SFN treatment could cause changes in cells that continue to impact HIV infection long after treatment. To address this possibility we tested the durability of the SFN-mediated anti-viral state in THP1-derived macrophages. All replicate cultures were treated with SFN for 24 hours. The SFN-containing media was then replaced with SFN-free media and new sets of cultures were exposed to freshly thawed aliquots of the same virus stock at 24-hour intervals. Each set of cultures was harvested for luciferase assays 24 hours after infection (Fig 3A). We found that infectability recovered over time in a dose-dependent manner (Fig 3B) with higher doses of SFN yielding a longer-lasting infection block. After removal from 10 μM SFN, cultures were as infectable as mock-treated controls in about 24 hours. These data show that the anti-viral effect induced by SFN treatment is reversible and that the SFN dose determines the duration over which the anti-viral state is maintained.


Sulforaphane Inhibits HIV Infection of Macrophages through Nrf2.

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

The SFN-mediated HIV infection block is reversible.(A), Study design: PMA-differentiated THP1 cells were pretreated with media supplemented with vehicle only (DMSO) or with 10 μM, 20 μM or 30 μM SFN. After twenty-four hours of treatment, the culture media was replaced with fresh SFN-free media. Cultures were infected 1, 2, 3, 4, or 5 days after treatment with freshly thawed aliquots of VSV-G pseudotyped HIV-1 encoding firefly luciferase in place of nef. Twenty-four hours after the start of each time-point infection, the cells were harvested and (B), luciferase activity was measured by photon emission. The bar graph represents the data for replicate experiments (n = 3).
© Copyright Policy
Related In: Results  -  Collection

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

ppat.1005581.g003: The SFN-mediated HIV infection block is reversible.(A), Study design: PMA-differentiated THP1 cells were pretreated with media supplemented with vehicle only (DMSO) or with 10 μM, 20 μM or 30 μM SFN. After twenty-four hours of treatment, the culture media was replaced with fresh SFN-free media. Cultures were infected 1, 2, 3, 4, or 5 days after treatment with freshly thawed aliquots of VSV-G pseudotyped HIV-1 encoding firefly luciferase in place of nef. Twenty-four hours after the start of each time-point infection, the cells were harvested and (B), luciferase activity was measured by photon emission. The bar graph represents the data for replicate experiments (n = 3).
Mentions: SFN treatment could cause changes in cells that continue to impact HIV infection long after treatment. To address this possibility we tested the durability of the SFN-mediated anti-viral state in THP1-derived macrophages. All replicate cultures were treated with SFN for 24 hours. The SFN-containing media was then replaced with SFN-free media and new sets of cultures were exposed to freshly thawed aliquots of the same virus stock at 24-hour intervals. Each set of cultures was harvested for luciferase assays 24 hours after infection (Fig 3A). We found that infectability recovered over time in a dose-dependent manner (Fig 3B) with higher doses of SFN yielding a longer-lasting infection block. After removal from 10 μM SFN, cultures were as infectable as mock-treated controls in about 24 hours. These data show that the anti-viral effect induced by SFN treatment is reversible and that the SFN dose determines the duration over which the anti-viral state is maintained.

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