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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 blocks HIV infection in monocytoid cell lines but does not increase SAMHD1 expression.(A), HeLa, (B), GHOST, (C), HEK293T, (D), PMA-differentiated U937, and (E), PMA-differentiated THP1, cells were treated with media supplemented with vehicle only (DMSO) or with 10 μM, 20 μM or 30 μM SFN. Pretreatment of cell cultures with 5 μM AZT served as a positive control for viral inhibition. Twenty-four hours after treatment, the samples were infected with VSV-G pseudotyped HIV-1 encoding firefly luciferase in place of nef. Twenty-four hours after infection, the cultures were harvested and luciferase activity was measured by photon emission. (F), PMA-differentiated THP1 cells were treated with SFN that underwent a twofold serial dilution with 10μM of SFN being the highest concentration. Twenty-four hours after treatment, the samples were either mock infected or infected with VSV-G pseudotyped HIV-1 encoding firefly luciferase in place of nef. Twenty-four hours after infection, luciferase activity was measured. The bar graphs represent the data for replicate experiments (n = 3). (G), SAMHD1 protein was detected by western blot analysis from lysates of mock- and SFN treated PMA-differentiated U937 cells. Lysates from mock-treated, PMA-differentiated THP1 cells served as a positive control for SAMHD1 production.
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ppat.1005581.g002: SFN blocks HIV infection in monocytoid cell lines but does not increase SAMHD1 expression.(A), HeLa, (B), GHOST, (C), HEK293T, (D), PMA-differentiated U937, and (E), PMA-differentiated THP1, cells were treated with media supplemented with vehicle only (DMSO) or with 10 μM, 20 μM or 30 μM SFN. Pretreatment of cell cultures with 5 μM AZT served as a positive control for viral inhibition. Twenty-four hours after treatment, the samples were infected with VSV-G pseudotyped HIV-1 encoding firefly luciferase in place of nef. Twenty-four hours after infection, the cultures were harvested and luciferase activity was measured by photon emission. (F), PMA-differentiated THP1 cells were treated with SFN that underwent a twofold serial dilution with 10μM of SFN being the highest concentration. Twenty-four hours after treatment, the samples were either mock infected or infected with VSV-G pseudotyped HIV-1 encoding firefly luciferase in place of nef. Twenty-four hours after infection, luciferase activity was measured. The bar graphs represent the data for replicate experiments (n = 3). (G), SAMHD1 protein was detected by western blot analysis from lysates of mock- and SFN treated PMA-differentiated U937 cells. Lysates from mock-treated, PMA-differentiated THP1 cells served as a positive control for SAMHD1 production.

Mentions: To gain a better understanding of the breadth of SFN anti-viral action, we tested HeLa, GHOST and HEK293T cells, as well as monocytoid U937 and THP1 cells that were differentiated into a macrophage-like state by incubation in PMA-supplemented media. The cells were pretreated for 24 hours, then infected and assayed as described above. We detected no inhibition of HIV infection after SFN treatment in HeLa, HEK293T or GHOST cells (Fig 2A–2C) but SFN markedly decreased evidence of infection in U937 and THP1 cultures (Fig 2D and 2E). Further titration of SFN in cultures of PMA-differentiated THP1 cells shows that infections can be reduced by 50% when cultures are treated with SFN in the 2.5 to 5 μM range (Fig 2F).


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 blocks HIV infection in monocytoid cell lines but does not increase SAMHD1 expression.(A), HeLa, (B), GHOST, (C), HEK293T, (D), PMA-differentiated U937, and (E), PMA-differentiated THP1, cells were treated with media supplemented with vehicle only (DMSO) or with 10 μM, 20 μM or 30 μM SFN. Pretreatment of cell cultures with 5 μM AZT served as a positive control for viral inhibition. Twenty-four hours after treatment, the samples were infected with VSV-G pseudotyped HIV-1 encoding firefly luciferase in place of nef. Twenty-four hours after infection, the cultures were harvested and luciferase activity was measured by photon emission. (F), PMA-differentiated THP1 cells were treated with SFN that underwent a twofold serial dilution with 10μM of SFN being the highest concentration. Twenty-four hours after treatment, the samples were either mock infected or infected with VSV-G pseudotyped HIV-1 encoding firefly luciferase in place of nef. Twenty-four hours after infection, luciferase activity was measured. The bar graphs represent the data for replicate experiments (n = 3). (G), SAMHD1 protein was detected by western blot analysis from lysates of mock- and SFN treated PMA-differentiated U937 cells. Lysates from mock-treated, PMA-differentiated THP1 cells served as a positive control for SAMHD1 production.
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ppat.1005581.g002: SFN blocks HIV infection in monocytoid cell lines but does not increase SAMHD1 expression.(A), HeLa, (B), GHOST, (C), HEK293T, (D), PMA-differentiated U937, and (E), PMA-differentiated THP1, cells were treated with media supplemented with vehicle only (DMSO) or with 10 μM, 20 μM or 30 μM SFN. Pretreatment of cell cultures with 5 μM AZT served as a positive control for viral inhibition. Twenty-four hours after treatment, the samples were infected with VSV-G pseudotyped HIV-1 encoding firefly luciferase in place of nef. Twenty-four hours after infection, the cultures were harvested and luciferase activity was measured by photon emission. (F), PMA-differentiated THP1 cells were treated with SFN that underwent a twofold serial dilution with 10μM of SFN being the highest concentration. Twenty-four hours after treatment, the samples were either mock infected or infected with VSV-G pseudotyped HIV-1 encoding firefly luciferase in place of nef. Twenty-four hours after infection, luciferase activity was measured. The bar graphs represent the data for replicate experiments (n = 3). (G), SAMHD1 protein was detected by western blot analysis from lysates of mock- and SFN treated PMA-differentiated U937 cells. Lysates from mock-treated, PMA-differentiated THP1 cells served as a positive control for SAMHD1 production.
Mentions: To gain a better understanding of the breadth of SFN anti-viral action, we tested HeLa, GHOST and HEK293T cells, as well as monocytoid U937 and THP1 cells that were differentiated into a macrophage-like state by incubation in PMA-supplemented media. The cells were pretreated for 24 hours, then infected and assayed as described above. We detected no inhibition of HIV infection after SFN treatment in HeLa, HEK293T or GHOST cells (Fig 2A–2C) but SFN markedly decreased evidence of infection in U937 and THP1 cultures (Fig 2D and 2E). Further titration of SFN in cultures of PMA-differentiated THP1 cells shows that infections can be reduced by 50% when cultures are treated with SFN in the 2.5 to 5 μM range (Fig 2F).

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