{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,5]],"date-time":"2026-09-05T04:32:21Z","timestamp":1788582741844,"version":"build-2803163510"},"reference-count":48,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2025,5,22]],"date-time":"2025-05-22T00:00:00Z","timestamp":1747872000000},"content-version":"vor","delay-in-days":21,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Medical Virology"],"published-print":{"date-parts":[[2025,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Hepatitis B e antigen (HBeAg) seroconversion is a crucial event in the natural history of chronic hepatitis B virus (HBV) infection, marked by a significant decrease in viral load and the emergence of mutations that suppress HBeAg expression. However, these mutations alone do not fully account for the reduction in viral load. This study investigated the biological features and pathogenic roles of mutations outside the basal core promoter (BCP) and precore regions during the HBeAg\u2010negative stage of chronic infection. Full\u2010length HBV genomes from HBeAg\u2010positive (<jats:italic>n<\/jats:italic>\u2009=\u2009180) and HBeAg\u2010negative (<jats:italic>n<\/jats:italic>\u2009=\u2009328) genotype D datasets were analyzed, revealing significantly higher genomic heterogeneity in HBeAg\u2010negative sequences compared with HBeAg\u2010positive genomes (50.4\u2009\u00b1\u200916.0 vs. 26.6\u2009\u00b1\u200910.5 nucleotide changes per genome). Twenty\u2010six hotspot amino acid mutations associated with the HBeAg\u2010negative stage were identified, with over half located in the Core region. Subsequently, full\u2010length HBV genomes from six HBeAg\u2010negative patient\u2010derived serum samples were obtained by PCR amplification followed by Sanger sequencing. Infectious clones generated from these genomes, each carrying between 21 and 66 amino acid substitutions, were characterized, showing that mutations in this stage differentially affected viral fitness in vitro by up\u2010 or downregulating HBV\u2010DNA levels (ranging from 0.2 to 5 times those of the wild\u2010type isolate), modulating capsid assembly, and altering the expression, secretion, and subcellular localization of viral proteins. In conclusion, while mutations in the BCP and precore regions are the primary drivers of HBeAg seroconversion, mutations outside these regions significantly influence HBV biology and potentially contribute to viral pathogenicity, underscoring the complex interplay between host and virus during the HBeAg\u2010negative stage of chronic infection.<\/jats:p>","DOI":"10.1002\/jmv.70398","type":"journal-article","created":{"date-parts":[[2025,5,22]],"date-time":"2025-05-22T03:23:57Z","timestamp":1747884237000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Unraveling the Role of Mutations Outside the Basal Promoter and Precore Regions in the HBeAg\u2010Negative Stage of Chronic Hepatitis B"],"prefix":"10.1002","volume":"97","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0533-4283","authenticated-orcid":false,"given":"Mar\u00eda Mercedes","family":"Elizalde","sequence":"first","affiliation":[{"name":"Universidad de Buenos Aires, Facultad de Medicina, Instituto de Investigaciones Biom\u00e9dicas en Retrovirus y Sida (INBIRS), CONICET Buenos Aires Argentina"},{"name":"Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas (CONICET) Buenos Aires Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mar\u00eda Cecilia","family":"Monzani","sequence":"additional","affiliation":[{"name":"Universidad de Buenos Aires, Facultad de Medicina, Instituto de Investigaciones Biom\u00e9dicas en Retrovirus y Sida (INBIRS), CONICET Buenos Aires Argentina"},{"name":"Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas (CONICET) Buenos Aires Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicol\u00e1s Octavio","family":"Favale","sequence":"additional","affiliation":[{"name":"Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas (CONICET) Buenos Aires Argentina"},{"name":"Universidad de Buenos Aires, Facultad de Farmacia y Bioqu\u00edmica, C\u00e1tedra de Biolog\u00eda Celular y Molecular, CONICET Buenos Aires Argentina"},{"name":"Universidad de Buenos Aires, Facultad de Farmacia y Bioqu\u00edmica, Instituto de Qu\u00edmica y Fisicoqu\u00edmica Biol\u00f3gicas \u201cProfesor Dr. Alejandro C. Paladini\u201d (IQUIFIB) Buenos Aires Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bel\u00e9n","family":"Bouzas","sequence":"additional","affiliation":[{"name":"Unidad de Virolog\u00eda, Hospital de Infecciosas \u201cFrancisco J. Mu\u00f1iz\u201d Buenos Aires Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lilia","family":"Mammana","sequence":"additional","affiliation":[{"name":"Unidad de Virolog\u00eda, Hospital de Infecciosas \u201cFrancisco J. 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