Persistent Loss of Hepatitis B Virus Markers in Serum without Cellular Immunity by Combination of Peginterferon and Entecavir Therapy in Humanized Mice

Antimicrob Agents Chemother. 2017 Aug 24;61(9):e00725-17. doi: 10.1128/AAC.00725-17. Print 2017 Sep.

Abstract

Nucleot(s)ide analogues and peginterferon (PEG-IFN) treatment are the only approved therapies for chronic hepatitis B virus (HBV) infection. However, complete eradication of the virus, as indicated by persistent loss of hepatitis B surface antigen (HBsAg), is rare among treated patients. This is due to long-term persistence of the HBV genome in infected hepatocytes in the form of covalently closed circular DNA (cccDNA). In this study, we investigated whether administration of a large dose of a nucleoside analogue in combination with PEG-IFN can achieve long-term loss of HBsAg in human hepatocyte chimeric mice. Mice were treated with a high dose of entecavir and/or PEG-IFN for 6 weeks. High-dose combination therapy with both drugs resulted in persistently negative HBV DNA in serum. Although small amounts of HBV DNA and cccDNA (0.1 and 0.01 copy/cell, respectively) remained in the mouse livers, some of the mice remained persistently negative for serum HBV DNA at 13 weeks after cessation of the therapy. Serum HBsAg and hepatitis B core-related antigen (HBcrAg) continued to decrease and eventually became negative at 12 weeks after cessation of the therapy. Analysis of the HBV genome in treated mice showed accumulation of G-to-A hypermutation and CpG III island methylation. Persistent loss of serum HBV DNA and loss of HBV markers by high-dose entecavir and PEG-IFN combination treatment in chimeric mice suggests that control of HBV can be achieved even in the absence of a cellular immune response.

Keywords: DNA methylation; cccDNA; hepatitis B virus; human hepatocyte chimeric mouse; hypermutation; methylation.

MeSH terms

  • Animals
  • Antiviral Agents / therapeutic use*
  • DNA Methylation / genetics
  • DNA, Circular / blood*
  • DNA, Viral / blood*
  • Drug Therapy, Combination
  • Guanine / analogs & derivatives*
  • Guanine / therapeutic use
  • Hepatitis B Surface Antigens / blood*
  • Hepatitis B virus / drug effects*
  • Hepatitis B virus / genetics
  • Hepatitis B virus / immunology
  • Hepatitis B, Chronic / drug therapy*
  • Hepatocytes / virology
  • Humans
  • Immunity, Cellular / immunology
  • Interferon-alpha / therapeutic use*
  • Liver / virology
  • Mice
  • Polyethylene Glycols / therapeutic use*
  • Recombinant Proteins / therapeutic use

Substances

  • Antiviral Agents
  • DNA, Circular
  • DNA, Viral
  • Hepatitis B Surface Antigens
  • Interferon-alpha
  • Recombinant Proteins
  • Polyethylene Glycols
  • entecavir
  • Guanine
  • peginterferon alfa-2a