{
  "abstract": "Background Hepatitis B surface antigen (HBsAg) can be derived from intrahepatic covalently closed circular DNA (cccDNA) and integrated hepatitis B virus (HBV) DNA (iDNA).Objective We evaluated the cccDNA and iDNA from liver tissues of 24 hepatitis B e antigen (HBeAg)(+) and 32 HBeAg(−) treatment-naïve chronic hepatitis B (CHB) participants in the North American Hepatitis B Research Network.Design For cccDNA analysis, DNA was heat-denatured and digested by plasmid-safe ATP-dependent DNase to remove relaxed circular DNA and iDNA before real-time polymerase chain reaction. For iDNA detection, total DNA was subjected to HBV hybridisation-targeted next generation sequencing assay for identification of the HBV-host junction sequences. Comparisons of HBV cccDNA and iDNA with other virological biomarkers were assessed.Results Intrahepatic cccDNA, serum HBV DNA, HBV RNA, hepatitis B core related antigen and quantitative hepatitis B surface antigen were higher in HBeAg(+) CHB. Intrahepatic hepatitis B core antigen staining was present in 87% HBeAg(+) but only 13% HBeAg(−) samples (p<0.0001). HBsAg staining was frequent in over 85% in both groups. 23 (95.8%) HBeAg(+) participants had ≤50% iDNA whereas 25 (78.1%) HBeAg(−) participants had >50% iDNA of total HBV DNA in their livers. For HBeAg(+) CHB, the iDNA integration sites were random with only 15.9% localised to the direct repeat 2 (DR2)-DR1 region. For HBeAg(−) CHB, 52.4% of the iDNA integrations were clustered at DR2-DR1. Microhomology-mediated end joining patterns of double-stranded linear DNA HBV integration was more frequent in HBeAg(+) livers.Conclusion HBeAg(−) CHB was associated with high HBsAg staining concentration despite low cccDNA levels suggesting that iDNA was the primary source of HBsAg. The high frequency of DR2-DR1 iDNA distribution in HBeAg(−) CHB suggests the selection advantage and clonal expansion of this integrant in the natural history of CHB.",
  "authors": [
    {
      "affiliations": [
        "Liver Center, Division of Gastroenterology and Hepatology, Beth Israel Deaconess Medical Center, Boston, Massachusetts, USA"
      ],
      "name": "Daryl T-Y Lau"
    },
    {
      "affiliations": [
        "Department of Microbiology and Molecular Genetics",
        "Cancer Virology Program, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA"
      ],
      "name": "Elena S Kim"
    },
    {
      "affiliations": [
        "JBS Science Inc, Doylestown, Pennsylvania, USA"
      ],
      "name": "Zhili Wang"
    },
    {
      "affiliations": [
        "Epidemiology, University of Pittsburgh Graduate School of Public Health, Pittsburgh, Pennsylvania, USA"
      ],
      "name": "Wendy C King"
    },
    {
      "affiliations": [
        "Laboratory of Pathology, The National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA"
      ],
      "name": "David E Kleiner"
    },
    {
      "affiliations": [
        "Liver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA"
      ],
      "name": "Marc G Ghany"
    },
    {
      "affiliations": [
        "Department of Microbiology and Molecular Genetics",
        "Cancer Virology Program, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA"
      ],
      "name": "Yuanjie Liu"
    },
    {
      "affiliations": [
        "Gastrointestinal Unit, Massachusetts General Hospital, Boston, Massachusetts, USA"
      ],
      "name": "Raymond Chung"
    },
    {
      "affiliations": [
        "Central Virginia Veterans Healthcare System, Richmond, Virginia, USA"
      ],
      "name": "Richard K Sterling"
    },
    {
      "affiliations": [
        "Infectious Disease Research, Abbott Laboratories, Abbott Park, Illinois, USA"
      ],
      "name": "Gavin Cloherty"
    },
    {
      "affiliations": [
        "JBS Science Inc, Doylestown, Pennsylvania, USA"
      ],
      "name": "Selena Y Lin"
    },
    {
      "affiliations": [
        "Translational Medical Science, The Baruch S. Blumberg Institute, Doylestown, Pennsylvania, USA"
      ],
      "name": "Hsin-Ni Liu"
    },
    {
      "affiliations": [
        "Department of Microbiology and Molecular Genetics",
        "Cancer Virology Program, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA"
      ],
      "name": "Ning Sun"
    },
    {
      "affiliations": [
        "JBS Science Inc, Doylestown, Pennsylvania, USA",
        "Translational Medical Science, The Baruch S. Blumberg Institute, Doylestown, Pennsylvania, USA"
      ],
      "name": "Ying-Hsiu Su"
    },
    {
      "affiliations": [
        "Department of Microbiology and Molecular Genetics",
        "Cancer Virology Program, UPMC Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA"
      ],
      "name": "Haitao Guo"
    }
  ],
  "title": "Differential intrahepatic integrated HBV DNA patterns between HBeAg-positive and HBeAg-negative chronic hepatitis B",
  "uid": "e4f44f87-4035-5b72-8dad-e8a318371b87"
}