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Interleukin-1 and tumor necrosis factor-α trigger restriction of hepatitis B virus infection via a cytidine deaminase activation-induced cytidine deaminase (AID).

Watashi K, Liang G, Iwamoto M, Marusawa H, Uchida N, Daito T, Kitamura K, Muramatsu M, Ohashi H, Kiyohara T, Suzuki R, Li J, Tong S, Tanaka Y, Murata K, Aizaki H, Wakita T - J. Biol. Chem. (2013)

Bottom Line: This effect was mediated by activation of the NF-κB signaling pathway.Although AID induced hypermutation of HBV DNA, this activity was dispensable for the anti-HBV activity.The antiviral effect of IL-1/TNFα was also observed on different HBV genotypes but not on hepatitis C virus.

View Article: PubMed Central - PubMed

Affiliation: From the Department of Virology II, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.

ABSTRACT
Virus infection is restricted by intracellular immune responses in host cells, and this is typically modulated by stimulation of cytokines. The cytokines and host factors that determine the host cell restriction against hepatitis B virus (HBV) infection are not well understood. We screened 36 cytokines and chemokines to determine which were able to reduce the susceptibility of HepaRG cells to HBV infection. Here, we found that pretreatment with IL-1β and TNFα remarkably reduced the host cell susceptibility to HBV infection. This effect was mediated by activation of the NF-κB signaling pathway. A cytidine deaminase, activation-induced cytidine deaminase (AID), was up-regulated by both IL-1β and TNFα in a variety of hepatocyte cell lines and primary human hepatocytes. Another deaminase APOBEC3G was not induced by these proinflammatory cytokines. Knockdown of AID expression impaired the anti-HBV effect of IL-1β, and overexpression of AID antagonized HBV infection, suggesting that AID was one of the responsible factors for the anti-HBV activity of IL-1/TNFα. Although AID induced hypermutation of HBV DNA, this activity was dispensable for the anti-HBV activity. The antiviral effect of IL-1/TNFα was also observed on different HBV genotypes but not on hepatitis C virus. These results demonstrate that proinflammatory cytokines IL-1/TNFα trigger a novel antiviral mechanism involving AID to regulate host cell permissiveness to HBV infection.

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NF-κB activation triggered by IL-1 and TNFα was critical for anti-HBV activity.A–D, F, and I, HepaRG cells were left untreated (control) or treated with varying concentrations of IL-1β (1, 10, 30, and 100 ng/ml) or 25 units/ml heparin (A), with 30 ng/ml IL-1β together with or without a neutralizing anti-IL-1RI antibody at 20 μg/ml (B), with 10 ng/ml IL-1β or varying concentrations of IL-1Ra (10, 30, and 100 ng/ml) (C), with 3 ng/ml IL-1β together with or without PD98059, SP600125, SB203580, or Bay11-7082 (D), or QNZ or BMS-345541 (F), or with TNFα (10, 100, and 300 ng/ml) (I) according to the treatment schedule shown in Fig. 1A. HBV infection was monitored by HBs protein secretion into the medium in A, C, D, F, and I and with HBc protein in the cells in B. E, G, and H, NF-κB (E and G) and ISRE activity (H) were measured by reporter assay in the cells transfected with the reporter plasmid expressing luciferase driven from five tandem repeats of NF-κB elements (E, upper graph, and G) or ISRE (H, upper graph) or by RT-PCR in the cells (E and H, lower panels) upon signaling inhibitors used in D and F together with or without IL-1β (E and G), or upon IL-1β (10, 30, and 100 ng/ml) or IFNα 100 IU/ml as a positive control (H) for 6 h. The white and black bars in the upper graph of E and G show the data in the absence or presence of IL-1β, respectively. Bands for mRNA for cIAP, XIAP, and GAPDH (E) or ISG56, PKR, or GAPDH (H) are presented in the lower panels. All of the data are based on averages of three independent experiments.
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Figure 2: NF-κB activation triggered by IL-1 and TNFα was critical for anti-HBV activity.A–D, F, and I, HepaRG cells were left untreated (control) or treated with varying concentrations of IL-1β (1, 10, 30, and 100 ng/ml) or 25 units/ml heparin (A), with 30 ng/ml IL-1β together with or without a neutralizing anti-IL-1RI antibody at 20 μg/ml (B), with 10 ng/ml IL-1β or varying concentrations of IL-1Ra (10, 30, and 100 ng/ml) (C), with 3 ng/ml IL-1β together with or without PD98059, SP600125, SB203580, or Bay11-7082 (D), or QNZ or BMS-345541 (F), or with TNFα (10, 100, and 300 ng/ml) (I) according to the treatment schedule shown in Fig. 1A. HBV infection was monitored by HBs protein secretion into the medium in A, C, D, F, and I and with HBc protein in the cells in B. E, G, and H, NF-κB (E and G) and ISRE activity (H) were measured by reporter assay in the cells transfected with the reporter plasmid expressing luciferase driven from five tandem repeats of NF-κB elements (E, upper graph, and G) or ISRE (H, upper graph) or by RT-PCR in the cells (E and H, lower panels) upon signaling inhibitors used in D and F together with or without IL-1β (E and G), or upon IL-1β (10, 30, and 100 ng/ml) or IFNα 100 IU/ml as a positive control (H) for 6 h. The white and black bars in the upper graph of E and G show the data in the absence or presence of IL-1β, respectively. Bands for mRNA for cIAP, XIAP, and GAPDH (E) or ISG56, PKR, or GAPDH (H) are presented in the lower panels. All of the data are based on averages of three independent experiments.

Mentions: As shown in Fig. 2A, IL-1β suppressed HBV infection in a dose-dependent manner. This anti-HBV effect was reversed by cotreatment with a neutralizing antibody for the IL-1 receptor, IL-1RI (Fig. 2B), suggesting that receptor engagement was required for anti-HBV activity. IL-1Ra is a natural antagonist that associates with IL-1RI but does not trigger downstream signal transduction (56). Treatment with IL-1Ra did not decrease HBV infectivity (Fig. 2C), suggesting that signal transduction triggered by IL-1 was required for anti-HBV activity.


Interleukin-1 and tumor necrosis factor-α trigger restriction of hepatitis B virus infection via a cytidine deaminase activation-induced cytidine deaminase (AID).

Watashi K, Liang G, Iwamoto M, Marusawa H, Uchida N, Daito T, Kitamura K, Muramatsu M, Ohashi H, Kiyohara T, Suzuki R, Li J, Tong S, Tanaka Y, Murata K, Aizaki H, Wakita T - J. Biol. Chem. (2013)

NF-κB activation triggered by IL-1 and TNFα was critical for anti-HBV activity.A–D, F, and I, HepaRG cells were left untreated (control) or treated with varying concentrations of IL-1β (1, 10, 30, and 100 ng/ml) or 25 units/ml heparin (A), with 30 ng/ml IL-1β together with or without a neutralizing anti-IL-1RI antibody at 20 μg/ml (B), with 10 ng/ml IL-1β or varying concentrations of IL-1Ra (10, 30, and 100 ng/ml) (C), with 3 ng/ml IL-1β together with or without PD98059, SP600125, SB203580, or Bay11-7082 (D), or QNZ or BMS-345541 (F), or with TNFα (10, 100, and 300 ng/ml) (I) according to the treatment schedule shown in Fig. 1A. HBV infection was monitored by HBs protein secretion into the medium in A, C, D, F, and I and with HBc protein in the cells in B. E, G, and H, NF-κB (E and G) and ISRE activity (H) were measured by reporter assay in the cells transfected with the reporter plasmid expressing luciferase driven from five tandem repeats of NF-κB elements (E, upper graph, and G) or ISRE (H, upper graph) or by RT-PCR in the cells (E and H, lower panels) upon signaling inhibitors used in D and F together with or without IL-1β (E and G), or upon IL-1β (10, 30, and 100 ng/ml) or IFNα 100 IU/ml as a positive control (H) for 6 h. The white and black bars in the upper graph of E and G show the data in the absence or presence of IL-1β, respectively. Bands for mRNA for cIAP, XIAP, and GAPDH (E) or ISG56, PKR, or GAPDH (H) are presented in the lower panels. All of the data are based on averages of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Figure 2: NF-κB activation triggered by IL-1 and TNFα was critical for anti-HBV activity.A–D, F, and I, HepaRG cells were left untreated (control) or treated with varying concentrations of IL-1β (1, 10, 30, and 100 ng/ml) or 25 units/ml heparin (A), with 30 ng/ml IL-1β together with or without a neutralizing anti-IL-1RI antibody at 20 μg/ml (B), with 10 ng/ml IL-1β or varying concentrations of IL-1Ra (10, 30, and 100 ng/ml) (C), with 3 ng/ml IL-1β together with or without PD98059, SP600125, SB203580, or Bay11-7082 (D), or QNZ or BMS-345541 (F), or with TNFα (10, 100, and 300 ng/ml) (I) according to the treatment schedule shown in Fig. 1A. HBV infection was monitored by HBs protein secretion into the medium in A, C, D, F, and I and with HBc protein in the cells in B. E, G, and H, NF-κB (E and G) and ISRE activity (H) were measured by reporter assay in the cells transfected with the reporter plasmid expressing luciferase driven from five tandem repeats of NF-κB elements (E, upper graph, and G) or ISRE (H, upper graph) or by RT-PCR in the cells (E and H, lower panels) upon signaling inhibitors used in D and F together with or without IL-1β (E and G), or upon IL-1β (10, 30, and 100 ng/ml) or IFNα 100 IU/ml as a positive control (H) for 6 h. The white and black bars in the upper graph of E and G show the data in the absence or presence of IL-1β, respectively. Bands for mRNA for cIAP, XIAP, and GAPDH (E) or ISG56, PKR, or GAPDH (H) are presented in the lower panels. All of the data are based on averages of three independent experiments.
Mentions: As shown in Fig. 2A, IL-1β suppressed HBV infection in a dose-dependent manner. This anti-HBV effect was reversed by cotreatment with a neutralizing antibody for the IL-1 receptor, IL-1RI (Fig. 2B), suggesting that receptor engagement was required for anti-HBV activity. IL-1Ra is a natural antagonist that associates with IL-1RI but does not trigger downstream signal transduction (56). Treatment with IL-1Ra did not decrease HBV infectivity (Fig. 2C), suggesting that signal transduction triggered by IL-1 was required for anti-HBV activity.

Bottom Line: This effect was mediated by activation of the NF-κB signaling pathway.Although AID induced hypermutation of HBV DNA, this activity was dispensable for the anti-HBV activity.The antiviral effect of IL-1/TNFα was also observed on different HBV genotypes but not on hepatitis C virus.

View Article: PubMed Central - PubMed

Affiliation: From the Department of Virology II, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.

ABSTRACT
Virus infection is restricted by intracellular immune responses in host cells, and this is typically modulated by stimulation of cytokines. The cytokines and host factors that determine the host cell restriction against hepatitis B virus (HBV) infection are not well understood. We screened 36 cytokines and chemokines to determine which were able to reduce the susceptibility of HepaRG cells to HBV infection. Here, we found that pretreatment with IL-1β and TNFα remarkably reduced the host cell susceptibility to HBV infection. This effect was mediated by activation of the NF-κB signaling pathway. A cytidine deaminase, activation-induced cytidine deaminase (AID), was up-regulated by both IL-1β and TNFα in a variety of hepatocyte cell lines and primary human hepatocytes. Another deaminase APOBEC3G was not induced by these proinflammatory cytokines. Knockdown of AID expression impaired the anti-HBV effect of IL-1β, and overexpression of AID antagonized HBV infection, suggesting that AID was one of the responsible factors for the anti-HBV activity of IL-1/TNFα. Although AID induced hypermutation of HBV DNA, this activity was dispensable for the anti-HBV activity. The antiviral effect of IL-1/TNFα was also observed on different HBV genotypes but not on hepatitis C virus. These results demonstrate that proinflammatory cytokines IL-1/TNFα trigger a novel antiviral mechanism involving AID to regulate host cell permissiveness to HBV infection.

Show MeSH
Related in: MedlinePlus