Duck Hepatitis B Virus: An Invaluable Model System for HBV Infection

Nucleic acid polymers (NAPs) are an attractive treatment modality for chronic hepatitis B (CHB), with REP2139 and REP2165 having shown efficacy in CHB patients. A subset of patients achieve functional cure, whereas the others exhibit a moderate response or are non-responders. NAP efficacy has been difficult to recapitulate in animal models, with the duck hepatitis B virus (DHBV) model showing some promise but remaining underexplored for NAP efficacy testing. Here we report on an optimized in vivo DHBV duck model and explore several characteristics of NAP treatment. REP2139 was efficacious in reducing DHBV DNA and DHBsAg levels in approximately half of the treated ducks, whether administered intraperitoneally or subcutaneously. Intrahepatic or serum NAP concentrations did not correlate with efficacy, nor did the appearance of anti-DHBsAg antibodies. Furthermore, NAP efficacy was only observed in experimentally infected ducks, not in endogenously infected ducks (vertical transmission). REP2139 add-on to entecavir treatment induced a deeper and more sustained virological response compared to entecavir monotherapy. Destabilized REP2165 showed a different activity profile with a more homogenous antiviral response followed by a faster rebound. In conclusion, subcutaneous administration of NAPs in the DHBV duck model provides a useful tool for in vivo evaluation of NAPs. It recapitulates many aspects of this class of compound's efficacy in CHB patients, most notably the clear division between responders and non-responders.

Owing to its high similarity to human hepatitis B virus (HBV), duck HBV (DHBV) is often used as an essential model for HBV research. Although intergenotypic recombination of HBV is common, it remains unclear whether the intergenotypic recombination of human HBV is exactly the same as that of DHBV. In this study, 119 serum samples of duck and goose were collected from 51 farms (29 duck and 22 goose farms) in the central and eastern regions of China. A total of 22 strains isolated from the 22 DHBV positive flock were sequenced. Genome sequence alignment revealed that the duck- and goose-origin strains shared the highest and lowest similarities (99.7 and 90.52%, respectively). The complete genomes of these DHBV and 31 reference strains were analyzed using phylogenetic methods and classified into 3 clusters, which corresponded to the previously identified DHBV-I, DHBV-II, and DHBV-III branches. Recombination analyses of the 53 DHBV genomes indicated 2 major intergenotypic recombination events with high confidence values. These recombination events occurred between the genotypes of the Chinese isolates Y180813HB (Chinese branch [DHBV-Ⅰ]) and E170101AH (Chinese branch [DHBV-Ⅱ]) and the Western isolate DHBV-XY (Western branch [DHBV-Ⅲ]), resulting in the emergence of 2 Chinese recombinant isolates Y190303HN and Y170101HB. In addition, 40% (2/5) goose-origin and 58.8% (10/17) duck-origin DHBV in this study harbored the mutation site of G133E in preS, which promote the pathogenicity of DHBV. This is the first study to report on the genome analysis and recombination characterization of DHBV isolated from Chinese geese. Further, continuous investigation and molecular identification of DHBV should be conducted to attract researchers’ attention.

Hepatitis B virus (HBV) poses a major global health burden with 260 million people being chronically infected and 890,000 dying annually from complications in the course of the infection. HBV is a small enveloped virus with a reverse-transcribed DNA genome that infects hepatocytes and can cause acute and chronic infections of the liver. HBV is endemic in humans and apes representing the prototype member of the viral family Hepadnaviridae and can be divided into 10 genotypes. Hepadnaviruses have been found in all vertebrate classes and constitute an ancient viral family that descended from non-enveloped progenitors more than 360 million years ago. The de novo emergence of the envelope protein gene was accompanied with the liver-tropism and resulted in a tight virus-host association. The oldest HBV genomes so far have been isolated from human remains of the Bronze Age and the Neolithic (~7000 years before present). Despite the remarkable stability of the hepadnaviral genome over geological eras, HBV is able to rapidly evolve within an infected individual under pressure of the immune response or during antiviral treatment. Treatment with currently available antivirals blocking intracellular replication of HBV allows controlling of high viremia and improving liver health during long-term therapy of patients with chronic hepatitis B (CHB), but they are not sufficient to cure the disease. New therapy options that cover all HBV genotypes and emerging viral variants will have to be developed soon. In addition to the antiviral treatment of chronically infected patients, continued efforts to expand the global coverage of the currently available HBV vaccine will be one of the key factors for controlling the rising global spread of HBV. Certain improvements of the vaccine (e.g. inclusion of PreS domains) could counteract known problems such as low or no responsiveness of certain risk groups and waning anti-HBs titers leading to occult infections, especially with HBV genotypes E or F. But even with an optimal vaccine and a cure for hepatitis B, global eradication of HBV would be difficult to achieve because of an existing viral reservoir in primates and bats carrying closely related hepadnaviruses with zoonotic potential.

The liver acts as an important filter for the body and is thus exposed to a wide range of pathogens. In order to study a hepatotropic pathogen’s intricate life cycle as well as associated disease and pathogenesis, authentic culture systems are vital. As such, human induced pluripotent stem cell (iPSC)-derived hepatocyte-like cells have recently been proposed as an attractive alternative to conventionally used primary human hepatocytes and hepatoma cell lines.

Here, we summarize current knowledge on the biology of major liver pathogens, including hepatotropic viruses A–E as well as the malaria parasite Plasmodium. We discuss the available systems used to study them, remaining questions in the field, and how iPSC technology may help to solve them. We also summarize limitations of available iPSC-derived hepatic systems and recent efforts to overcome them. With their ability to provide a universally susceptible system for personalized, complex, and at the same time isogenic infection models, we believe that iPSCs have the potential to transform the future of hepatotropic infectious disease medicine.

Programmed death-1 (PD-1), upon engagement by its ligands, programmed death ligand-1 (PD-L1) and programmed death ligand-2 (PD-L2), provides signals that attenuate adaptive immune responses. Here we describe the identification of the Pekin duck PD-L2 (duPD-L2) and its gene structure. The duPD-L2 cDNA encodes a 321 amino acid protein that has an amino acid identity of 76% and 35% with chicken and human PD-L2, respectively. Mapping of the duPD-L2 cDNA with duck genomic sequences revealed an exonic structure similar to that of the human Pdcd1lg2 gene. Homology modelling of the duPD-L2 protein was compatible with the murine PD-L2 ectodomain structure. Residues known to be important for PD-1 receptor binding of murine PD-L2 were mostly conserved in duPD-L2 within sheets A and G and partially conserved within sheets C and F. DuPD-L2 mRNA was constitutively expressed in all tissues examined with highest expression levels in lung, spleen, cloaca, bursa, cecal tonsil, duodenum and very low levels of expression in muscle, kidney and brain. Lipopolysaccharide treatment of adherent duck PBMC upregulated duPD-L2 mRNA expression. Our work shows evolutionary conservation of the PD-L2 ectodomain structure and residues important for PD-1 binding in vertebrates including fish. The information provided will be useful for further investigation of the role of duPD-L2 in the regulation of duck adaptive immunity and exploration of PD-1-targeted immunotherapies in the duck hepatitis B infection model.

Globally, over half a billion people are persistently infected with hepatitis B (HBV) and/or hepatitis C viruses. Chronic HBV and HCV infection frequently lead to fibrosis, cirrhosis and hepatocellular carcinoma (HCC). Co-infections with hepatitis delta virus (HDV), a subviral satellite requiring HBV for its propagation, accelerates the progression of liver disease toward HCC. The mechanisms by which these viruses cause malignant transformation, culminating in HCC, remain incompletely understood, partially due to the lack of adequate experimental models for dissecting these complex disease processes in vivo.