Immunostimulatory siRNA with a uridine bulge leads to potent inhibition of HBV and activation of innate immunity
Abstract Background Hepatitis B virus (HBV) infection is difficult to cure. HBV-specific immune tolerance plays a key role in HBV persistence, and enhancing cellular and humoral immunity will improve the control of HBV infection. The purpose of the study was to explore the anti-HBV and immunostimula...
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| Format: | Article |
| Language: | English |
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BMC
2021-02-01
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| Series: | Virology Journal |
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| Online Access: | https://doi.org/10.1186/s12985-021-01509-z |
| _version_ | 1818325611023695872 |
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| author | Tingyu Lan Zhiqiang Wei Yulin He Song Wan Li Liu Bin Cheng Ruimin Li Hongxia Chen Guohua Liu Zhongji Meng |
| author_facet | Tingyu Lan Zhiqiang Wei Yulin He Song Wan Li Liu Bin Cheng Ruimin Li Hongxia Chen Guohua Liu Zhongji Meng |
| author_sort | Tingyu Lan |
| collection | DOAJ |
| description | Abstract Background Hepatitis B virus (HBV) infection is difficult to cure. HBV-specific immune tolerance plays a key role in HBV persistence, and enhancing cellular and humoral immunity will improve the control of HBV infection. The purpose of the study was to explore the anti-HBV and immunostimulatory effects of msiRNAs that introduce unpaired uridine bulges in the passenger strand. Methods msiRNAs targeting the HBV S and X genes were designed and named msiHBs and msiHBx, respectively. HepG2 cells were cotransfected with siRNA or msiRNA and the HBV replication-competent plasmid pHY106-wta or pHY106-X15. HepG2.215 cells were transfected with siRNA or msiRNA. The levels of HBsAg, HBeAg, and the cytokines TNF-α, IFN-α, IFN-β, IL-1α, and IL-6 in the culture supernatant was detected by ELISA. The levels of intracellular HBV RNA, nuclear HBV replication intermediates, and HBV DNA in the supernatant were measured by quantitative RT-PCR and PCR. The levels of HBV replication intermediates were detected by Southern blotting. Peripheral blood mononuclear cells were transfected with siRNA or msiRNA, and the levels of secreted cytokines IFN-α and IFN-β were detected by ELISA. The bioactivity of type I interferons in the supernatants was detected by the virus protection assay. Results msiHBx treatment led to a significant decrease in HBsAg (to a negative level) and HBV DNA (95.5%) in the supernatant and intrahepatocellular HBV replication intermediates (89.8%) in HepG2 cells with transient HBV replication and in HepG2.2.15 cells. There was no significant difference between msiHBx and siHBx in terms of the reduction in HBV proteins and HBV replication (P > 0.05). Compared with siHBx, msiHBx treatment of HepG2 cells transfected with the HBV replication-competent plasmid led to a significant increase in the levels of the antiviral cytokines TNF-α (3.3-fold), IFN-α (1.4-fold), and IFN-β (2.5-fold) (P < 0.01), without upregulation of the proinflammatory cytokines IL-1α and IL-6. The virus protection assay results showed msiHBx-mediated type I interferons effectively protected L929 cells against ECMV infection. Conclusions msiHBx could effectively inhibit HBV expression and replication and induce an antiviral innate immune response without proinflammatory activation. The dual RNAi and immunostimulatory activity of msiRNAs may play an important role in the control of HBV infection. |
| first_indexed | 2024-12-13T11:47:14Z |
| format | Article |
| id | doaj.art-5302d44df4164f6fa49abb11fe8905f0 |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-12-13T11:47:14Z |
| publishDate | 2021-02-01 |
| publisher | BMC |
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| series | Virology Journal |
| spelling | doaj.art-5302d44df4164f6fa49abb11fe8905f02022-12-21T23:47:28ZengBMCVirology Journal1743-422X2021-02-0118111010.1186/s12985-021-01509-zImmunostimulatory siRNA with a uridine bulge leads to potent inhibition of HBV and activation of innate immunityTingyu Lan0Zhiqiang Wei1Yulin He2Song Wan3Li Liu4Bin Cheng5Ruimin Li6Hongxia Chen7Guohua Liu8Zhongji Meng9Postgraduate Training Basement of Jinzhou Medicical University, Taihe Hospital, Hubei University of MedicineInstitute of Biomedical Research, Taihe Hospital, Hubei University of MedicineInstitute of Biomedical Research, Taihe Hospital, Hubei University of MedicineDepartment of Infectious Diseases, Taihe Hospital, Hubei University of MedicineDepartment of Infectious Diseases, Taihe Hospital, Hubei University of MedicineInstitute of Biomedical Research, Taihe Hospital, Hubei University of MedicineInstitute of Biomedical Research, Taihe Hospital, Hubei University of MedicineInstitute of Biomedical Research, Taihe Hospital, Hubei University of MedicineInstitute of Biomedical Research, Taihe Hospital, Hubei University of MedicinePostgraduate Training Basement of Jinzhou Medicical University, Taihe Hospital, Hubei University of MedicineAbstract Background Hepatitis B virus (HBV) infection is difficult to cure. HBV-specific immune tolerance plays a key role in HBV persistence, and enhancing cellular and humoral immunity will improve the control of HBV infection. The purpose of the study was to explore the anti-HBV and immunostimulatory effects of msiRNAs that introduce unpaired uridine bulges in the passenger strand. Methods msiRNAs targeting the HBV S and X genes were designed and named msiHBs and msiHBx, respectively. HepG2 cells were cotransfected with siRNA or msiRNA and the HBV replication-competent plasmid pHY106-wta or pHY106-X15. HepG2.215 cells were transfected with siRNA or msiRNA. The levels of HBsAg, HBeAg, and the cytokines TNF-α, IFN-α, IFN-β, IL-1α, and IL-6 in the culture supernatant was detected by ELISA. The levels of intracellular HBV RNA, nuclear HBV replication intermediates, and HBV DNA in the supernatant were measured by quantitative RT-PCR and PCR. The levels of HBV replication intermediates were detected by Southern blotting. Peripheral blood mononuclear cells were transfected with siRNA or msiRNA, and the levels of secreted cytokines IFN-α and IFN-β were detected by ELISA. The bioactivity of type I interferons in the supernatants was detected by the virus protection assay. Results msiHBx treatment led to a significant decrease in HBsAg (to a negative level) and HBV DNA (95.5%) in the supernatant and intrahepatocellular HBV replication intermediates (89.8%) in HepG2 cells with transient HBV replication and in HepG2.2.15 cells. There was no significant difference between msiHBx and siHBx in terms of the reduction in HBV proteins and HBV replication (P > 0.05). Compared with siHBx, msiHBx treatment of HepG2 cells transfected with the HBV replication-competent plasmid led to a significant increase in the levels of the antiviral cytokines TNF-α (3.3-fold), IFN-α (1.4-fold), and IFN-β (2.5-fold) (P < 0.01), without upregulation of the proinflammatory cytokines IL-1α and IL-6. The virus protection assay results showed msiHBx-mediated type I interferons effectively protected L929 cells against ECMV infection. Conclusions msiHBx could effectively inhibit HBV expression and replication and induce an antiviral innate immune response without proinflammatory activation. The dual RNAi and immunostimulatory activity of msiRNAs may play an important role in the control of HBV infection.https://doi.org/10.1186/s12985-021-01509-zHepatitis B virusRNAimicroRNA-like siRNAmsiRNAImmunostimulationInnate immunity |
| spellingShingle | Tingyu Lan Zhiqiang Wei Yulin He Song Wan Li Liu Bin Cheng Ruimin Li Hongxia Chen Guohua Liu Zhongji Meng Immunostimulatory siRNA with a uridine bulge leads to potent inhibition of HBV and activation of innate immunity Virology Journal Hepatitis B virus RNAi microRNA-like siRNA msiRNA Immunostimulation Innate immunity |
| title | Immunostimulatory siRNA with a uridine bulge leads to potent inhibition of HBV and activation of innate immunity |
| title_full | Immunostimulatory siRNA with a uridine bulge leads to potent inhibition of HBV and activation of innate immunity |
| title_fullStr | Immunostimulatory siRNA with a uridine bulge leads to potent inhibition of HBV and activation of innate immunity |
| title_full_unstemmed | Immunostimulatory siRNA with a uridine bulge leads to potent inhibition of HBV and activation of innate immunity |
| title_short | Immunostimulatory siRNA with a uridine bulge leads to potent inhibition of HBV and activation of innate immunity |
| title_sort | immunostimulatory sirna with a uridine bulge leads to potent inhibition of hbv and activation of innate immunity |
| topic | Hepatitis B virus RNAi microRNA-like siRNA msiRNA Immunostimulation Innate immunity |
| url | https://doi.org/10.1186/s12985-021-01509-z |
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