{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,19]],"date-time":"2026-02-19T01:21:07Z","timestamp":1771464067945,"version":"3.50.1"},"reference-count":32,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2025,1,3]],"date-time":"2025-01-03T00:00:00Z","timestamp":1735862400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIH","award":["P50AA030407-5129"],"award-info":[{"award-number":["P50AA030407-5129"]}]},{"name":"NIH","award":["R01AI163042"],"award-info":[{"award-number":["R01AI163042"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>About 296 million people worldwide are living with chronic hepatitis B viral (HBV) infection, and outcomes to end-stage liver diseases are potentiated by alcohol. HBV replicates in hepatocytes, but other liver non-parenchymal cells can sense the virus. In this study, we aimed to investigate the regulatory effects of macrophages on HBV marker and interferon-stimulated genes (ISGs) expressions in hepatocytes. This study was performed on HBV-replicating HepG2.2.15 cells and human monocyte-derived macrophages (MDMs). We found that exposure of HepG2.2.15 cells to an acetaldehyde-generating system (AGS) increased HBV RNA, HBV DNA, and cccDNA expressions and suppressed the activation of ISGs, APOBEC3G, ISG15, and OAS1. Supernatants collected from IFN\u03b1-activated MDMs decreased HBV marker levels and induced ISG activation in AGS-treated and untreated HepG2.215 cells. These effects were reversed by exposure of MDMs to ethanol and mimicked by treatment with exosome release inhibitor GW4869. We conclude that exosome-mediated crosstalk between IFN-activated macrophages and HBV-replicating hepatocytes plays a protective role via the up-regulation of ISGs and suppression of HBV replication. However, ethanol exposure to macrophages breaks this protection.<\/jats:p>","DOI":"10.3390\/biom15010057","type":"journal-article","created":{"date-parts":[[2025,1,3]],"date-time":"2025-01-03T07:10:17Z","timestamp":1735888217000},"page":"57","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Ethanol Disrupts the Protective Crosstalk Between Macrophages and HBV-Infected Hepatocytes"],"prefix":"10.3390","volume":"15","author":[{"given":"Murali","family":"Ganesan","sequence":"first","affiliation":[{"name":"Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE 68198, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1517-2919","authenticated-orcid":false,"given":"Anup S.","family":"Pathania","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE 68198, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Grace","family":"Bybee","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE 68198, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7759-8889","authenticated-orcid":false,"given":"Kusum K.","family":"Kharbanda","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE 68105, USA"},{"name":"Research Service, Veterans Affairs Nebraska Western Iowa Health Care System, Omaha, NE 68105, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7339-8732","authenticated-orcid":false,"given":"Larisa Y.","family":"Poluektova","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE 68198, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7498-0556","authenticated-orcid":false,"given":"Natalia A.","family":"Osna","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE 68198, USA"},{"name":"Department of Internal Medicine, University of Nebraska Medical Center, Omaha, NE 68105, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,1,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1429","DOI":"10.14309\/ajg.0000000000000651","article-title":"Prevalence of Chronic Hepatitis B Virus Infection in the United States","volume":"115","author":"Lim","year":"2020","journal-title":"Am. J. Gastroenterol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2651","DOI":"10.3748\/wjg.v23.i15.2651","article-title":"Hepatitis B virus infection and alcohol consumption","volume":"23","author":"Enomoto","year":"2017","journal-title":"World J. Gastroenterol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"10052","DOI":"10.12998\/wjcc.v9.i33.10052","article-title":"Effects of alcohol consumption on viral hepatitis B and C","volume":"9","author":"Xu","year":"2021","journal-title":"World J. Clin. Cases"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"104925","DOI":"10.1016\/j.antiviral.2020.104925","article-title":"Hepatitis B virus biology and life cycle","volume":"182","author":"Tsukuda","year":"2020","journal-title":"Antivir. Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1718","DOI":"10.1111\/liv.13884","article-title":"Liver macrophages: Friend or foe during hepatitis B infection?","volume":"38","author":"Durantel","year":"2018","journal-title":"Liver Int."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Wang, J., Lu, H., and Li, Q. (2024). Hepatic macrophage niche: A bridge between HBV-mediated metabolic changes with intrahepatic inflammation. Front. Immunol., 15.","DOI":"10.3389\/fimmu.2024.1414594"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1152","DOI":"10.3748\/wjg.v17.i9.1152","article-title":"Hepatoma cell line HepG2.2.15 demonstrates distinct biological features compared with parental HepG2","volume":"17","author":"Zhao","year":"2011","journal-title":"World J. Gastroenterol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"G127","DOI":"10.1152\/ajpgi.00064.2019","article-title":"Acetaldehyde suppresses the display of HBV-MHC class I complexes on HBV-expressing hepatocytes","volume":"317","author":"Ganesan","year":"2019","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"883","DOI":"10.3748\/wjg.v26.i9.883","article-title":"Role of alcohol in pathogenesis of hepatitis B virus infection","volume":"26","author":"Ganesan","year":"2020","journal-title":"World J. Gastroenterol."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"New-Aaron, M., Thomes, P.G., Ganesan, M., Dagur, R.S., Donohue, T.M., Kusum, K.K., Poluektova, L.Y., and Osna, N.A. (2021). Alcohol-Induced Lysosomal Damage and Suppression of Lysosome Biogenesis Contribute to Hepatotoxicity in HIV-Exposed Liver Cells. Biomolecules, 11.","DOI":"10.3390\/biom11101497"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"G566","DOI":"10.1152\/ajpgi.00183.2015","article-title":"Acetaldehyde accelerates HCV-induced impairment of innate immunity by suppressing methylation reactions in liver cells","volume":"309","author":"Ganesan","year":"2015","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1038\/nprot.2013.032","article-title":"Mouse model of chronic and binge ethanol feeding (the NIAAA model)","volume":"8","author":"Bertola","year":"2013","journal-title":"Nat. Protoc."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"731","DOI":"10.1016\/S0092-8674(01)00269-0","article-title":"Arginine Methylation of STAT1 Modulates IFN\u03b1\/\u03b2-Induced Transcription","volume":"104","author":"Mowen","year":"2001","journal-title":"Cell"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1016\/j.jcmgh.2017.10.004","article-title":"Demethylase JMJD6 as a New Regulator of Interferon Signaling: Effects of HCV and Ethanol Metabolism","volume":"5","author":"Ganesan","year":"2018","journal-title":"Cell. Mol. Gastroenterol. Hepatol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Zhang, Z., Liu, J., Yu, L., Zeng, R., and Pan, W. (2024). The hijacking of HBV by small extracellular vesicles inhibits M1 macrophages to facilitate immune evasion. Sci. Rep., 14.","DOI":"10.1038\/s41598-024-70924-3"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1779","DOI":"10.1002\/hep.29348","article-title":"Hepatitis B virus evades innate immunity of hepatocytes but activates cytokine production by macrophages","volume":"66","author":"Cheng","year":"2017","journal-title":"Hepatology"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1639","DOI":"10.1006\/cyto.2000.0766","article-title":"Role of Type I Interferons During Macrophage Activation by Lipopolysaccharide","volume":"12","author":"Vadiveloo","year":"2000","journal-title":"Cytokine"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Megahed, F.A.K., Zhou, X., and Sun, P. (2020). The Interactions Between HBV and the Innate Immunity of Hepatocytes. Viruses, 12.","DOI":"10.3390\/v12030285"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1086","DOI":"10.1016\/j.jhep.2019.06.032","article-title":"Hepatitis B virus-induced modulation of liver macrophage function promotes hepatocyte infection","volume":"71","author":"Delphin","year":"2019","journal-title":"J. Hepatol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Li, Q., Sun, B., Zhuo, Y., Jiang, Z., Li, R., Lin, C., Jin, Y., Gao, Y., and Wang, D. (2022). Interferon and interferon-stimulated genes in HBV treatment. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.1034968"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Kakizaki, M., Yamamoto, Y., Yabuta, S., Kurosaki, N., Kagawa, T., and Kotani, A. (2018). The immunological function of extracellular vesicles in hepatitis B virus-infected hepatocytes. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0205886"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1038\/cmi.2016.24","article-title":"Exosomes mediate hepatitis B virus (HBV) transmission and NK-cell dysfunction","volume":"14","author":"Yang","year":"2017","journal-title":"Cell. Mol. Immunol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"793","DOI":"10.1038\/ni.2647","article-title":"Exosomes mediate the cell-to-cell transmission of IFN-\u03b1-induced antiviral activity","volume":"14","author":"Li","year":"2013","journal-title":"Nat. Immunol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"424","DOI":"10.14218\/JCTH.2020.00046","article-title":"Macrophage Phenotypes and Hepatitis B Virus Infection","volume":"8","author":"Li","year":"2020","journal-title":"J. Clin. Transl. Hepatol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Malherbe, D.C., and Messaoudi, I. (2022). Transcriptional and Epigenetic Regulation of Monocyte and Macrophage Dysfunction by Chronic Alcohol Consumption. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.911951"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Sun, J., Zhao, P., Shi, Y., and Li, Y. (2023). Recent insight into the role of macrophage in alcohol-associated liver disease: A mini-review. Front. Cell Dev. Biol., 11.","DOI":"10.3389\/fcell.2023.1292016"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"415","DOI":"10.21037\/apm-20-2353","article-title":"Effect of alcohol on the progress of hepatitis B cirrhosis","volume":"10","author":"Zhou","year":"2021","journal-title":"Ann. Palliat. Med."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Tan, G., Song, H., Xu, F., and Cheng, G. (2018). When Hepatitis B Virus Meets Interferons. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.01611"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2074","DOI":"10.1080\/21655979.2021.1931640","article-title":"Roles of APOBEC3 in hepatitis B virus (HBV) infection and hepatocarcinogenesis","volume":"12","author":"Zhang","year":"2021","journal-title":"Bioengineered"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1394","DOI":"10.3390\/v2071394","article-title":"Innate Antiviral Immune Responses to Hepatitis B Virus","volume":"2","author":"Lucifora","year":"2010","journal-title":"Viruses"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"101969","DOI":"10.1016\/j.isci.2020.101969","article-title":"Suppression of hepatitis B virus through therapeutic activation of RIG-I and IRF3 signaling in hepatocytes","volume":"24","author":"Lee","year":"2021","journal-title":"iScience"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Osna, N.A., Eguchi, A., Feldstein, A.E., Tsukamoto, H., Dagur, R.S., Ganesan, M., New-Aaron, M., Arumugam, M.K., Chava, S., and Ribeiro, M. (2022). Cell-to-Cell Communications in Alcohol-Associated Liver Disease. Front. Physiol., 13.","DOI":"10.3389\/fphys.2022.831004"}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/15\/1\/57\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T10:22:13Z","timestamp":1759918933000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/15\/1\/57"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,3]]},"references-count":32,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,1]]}},"alternative-id":["biom15010057"],"URL":"https:\/\/doi.org\/10.3390\/biom15010057","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,1,3]]}}}