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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 action impacts HIV-2 as well as HIV-1 and is not reporter dependent.PMA-differentiated THP1 cells were treated with media supplemented with vehicle only (DMSO), 5 μM AZT or with 10 μM SFN. Twenty-four hours after treatment, the samples were either mock infected or infected with (A), VSV-G pseudotyped HIV-1 encoding firefly luciferase in place of nef or (B), VSV-G pseudotyped HIV-2 encoding firefly luciferase in place of nef. Twenty-four hours after infection, luciferase activity was measured by photon emission. (C), In parallel, the same experiment as in (A) and (B) was performed except that THP1 cells were infected with VSV-G-pseudotyped HIV-1 with GFP in place of nef or (D), VSV-G pseudotyped HIV-2 with GFP in place of nef. The samples with GFP-reporter viruses were fixed and harvested 24 h after infection and the fraction of GFP(+) cells was enumerated by flow cytometry. Bar graphs represent the data for replicate experiments (n = 3).
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ppat.1005581.g004: SFN action impacts HIV-2 as well as HIV-1 and is not reporter dependent.PMA-differentiated THP1 cells were treated with media supplemented with vehicle only (DMSO), 5 μM AZT or with 10 μM SFN. Twenty-four hours after treatment, the samples were either mock infected or infected with (A), VSV-G pseudotyped HIV-1 encoding firefly luciferase in place of nef or (B), VSV-G pseudotyped HIV-2 encoding firefly luciferase in place of nef. Twenty-four hours after infection, luciferase activity was measured by photon emission. (C), In parallel, the same experiment as in (A) and (B) was performed except that THP1 cells were infected with VSV-G-pseudotyped HIV-1 with GFP in place of nef or (D), VSV-G pseudotyped HIV-2 with GFP in place of nef. The samples with GFP-reporter viruses were fixed and harvested 24 h after infection and the fraction of GFP(+) cells was enumerated by flow cytometry. Bar graphs represent the data for replicate experiments (n = 3).

Mentions: Here we tested whether SFN also impairs HIV-2 infection and whether SFN action is reporter-dependent. Cells were mock treated, or pretreated with vehicle, 5 μM AZT or 10 μM SFN, as indicated, and infected with VSV-G-pseudotyped HIV-1 or HIV-2 encoding luciferase in place of nef or with VSV-G-pseudotyped HIV-1 or HIV-2 encoding GFP in place of nef. Luciferase activity was measured in culture lysates and the percentage of GFP(+) cells was determined using flow cytometry. The loss of luciferase signal after SFN treatment in HIV-2-infected cultures paralleled that in the HIV-1-infected cultures (Fig 4A versus 4B). The decrease in the percentage of GFP(+) cells upon AZT or SFN treatment was also the same in HIV-1 and HIV-2 infected cultures (Fig 4A and 4B versus 4C and 4D). These results demonstrate that our observations, showing an SFN-mediated infection block, are not reporter-dependent. The similarity between the results for HIV-1 and HIV-2 further indicate that SAMHD1 is not responsible for the block because HIV-2 Vpx would be expected to at least partially overcome that restriction. The results also support a model, based on our flow cytometry data with the GFP reporter virus, in which fewer cells are becoming infected after SFN treatment, rather than one in which a similar number of infected cells produce less reporter protein.


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 action impacts HIV-2 as well as HIV-1 and is not reporter dependent.PMA-differentiated THP1 cells were treated with media supplemented with vehicle only (DMSO), 5 μM AZT or with 10 μM SFN. Twenty-four hours after treatment, the samples were either mock infected or infected with (A), VSV-G pseudotyped HIV-1 encoding firefly luciferase in place of nef or (B), VSV-G pseudotyped HIV-2 encoding firefly luciferase in place of nef. Twenty-four hours after infection, luciferase activity was measured by photon emission. (C), In parallel, the same experiment as in (A) and (B) was performed except that THP1 cells were infected with VSV-G-pseudotyped HIV-1 with GFP in place of nef or (D), VSV-G pseudotyped HIV-2 with GFP in place of nef. The samples with GFP-reporter viruses were fixed and harvested 24 h after infection and the fraction of GFP(+) cells was enumerated by flow cytometry. Bar graphs represent 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.g004: SFN action impacts HIV-2 as well as HIV-1 and is not reporter dependent.PMA-differentiated THP1 cells were treated with media supplemented with vehicle only (DMSO), 5 μM AZT or with 10 μM SFN. Twenty-four hours after treatment, the samples were either mock infected or infected with (A), VSV-G pseudotyped HIV-1 encoding firefly luciferase in place of nef or (B), VSV-G pseudotyped HIV-2 encoding firefly luciferase in place of nef. Twenty-four hours after infection, luciferase activity was measured by photon emission. (C), In parallel, the same experiment as in (A) and (B) was performed except that THP1 cells were infected with VSV-G-pseudotyped HIV-1 with GFP in place of nef or (D), VSV-G pseudotyped HIV-2 with GFP in place of nef. The samples with GFP-reporter viruses were fixed and harvested 24 h after infection and the fraction of GFP(+) cells was enumerated by flow cytometry. Bar graphs represent the data for replicate experiments (n = 3).
Mentions: Here we tested whether SFN also impairs HIV-2 infection and whether SFN action is reporter-dependent. Cells were mock treated, or pretreated with vehicle, 5 μM AZT or 10 μM SFN, as indicated, and infected with VSV-G-pseudotyped HIV-1 or HIV-2 encoding luciferase in place of nef or with VSV-G-pseudotyped HIV-1 or HIV-2 encoding GFP in place of nef. Luciferase activity was measured in culture lysates and the percentage of GFP(+) cells was determined using flow cytometry. The loss of luciferase signal after SFN treatment in HIV-2-infected cultures paralleled that in the HIV-1-infected cultures (Fig 4A versus 4B). The decrease in the percentage of GFP(+) cells upon AZT or SFN treatment was also the same in HIV-1 and HIV-2 infected cultures (Fig 4A and 4B versus 4C and 4D). These results demonstrate that our observations, showing an SFN-mediated infection block, are not reporter-dependent. The similarity between the results for HIV-1 and HIV-2 further indicate that SAMHD1 is not responsible for the block because HIV-2 Vpx would be expected to at least partially overcome that restriction. The results also support a model, based on our flow cytometry data with the GFP reporter virus, in which fewer cells are becoming infected after SFN treatment, rather than one in which a similar number of infected cells produce less reporter protein.

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