Florian A Lempp1,2, Ellen Wiedtke3, Bingqian Qu1, Pierre Roques4,5, Isabelle Chemin6, Florian W R Vondran7,8, Roger Le Grand4,5, Dirk Grimm3, Stephan Urban1,2. 1. Department of Infectious Diseases, Molecular Virology, University Hospital Heidelberg, Heidelberg, Germany. 2. German Centre for Infection Research, partner site Heidelberg, Heidelberg, Germany. 3. Cluster of Excellence CellNetworks, Department of Infectious Diseases, Virology, BioQuant, University Hospital Heidelberg, Heidelberg, Germany. 4. Division of ImmunoVirology, Institute of Emerging Diseases and Innovative Therapies, Centre d'Energie Atomique, Fontenay aux Roses, Paris, France. 5. UMRE01, UMR1184, Université Paris Sud, Orsay, France. 6. Université de Lyon, INSERM U1052, CNRS UMR5286, Centre de Recherche en Cancérologie de Lyon, Lyon, France. 7. Regenerative Medicine and Experimental Surgery, Department of General, Visceral and Transplantation Surgery, Hannover Medical School, Hannover, Germany. 8. German Centre for Infection Research (DZIF), partner site Hannover-Braunschweig, Hannover, Germany.
Abstract
Infections with the human hepatitis B virus (HBV) and hepatitis D virus (HDV) depend on species-specific host factors like the receptor human sodium taurocholate cotransporting polypeptide (hNTCP). Complementation of mouse hepatocytes with hNTCP confers susceptibility to HDV but not HBV, indicating the requirement of additional HBV-specific factors. As an essential premise toward the establishment of an HBV-susceptible animal model, we investigated the role of hNTCP as a limiting factor of hepatocytes in commonly used laboratory animals. Primary hepatocytes from mice, rats, dogs, pigs, rhesus macaques, and cynomolgus macaques were transduced with adeno-associated viral vectors encoding hNTCP and subsequently infected with HBV. Cells were analyzed for Myrcludex B binding, taurocholate uptake, HBV covalently closed circular DNA formation, and expression of all HBV markers. Sodium taurocholate cotransporting polypeptide (Ntcp) from the respective species was cloned and analyzed for HBV and HDV receptor activity in a permissive hepatoma cell line. Expression of hNTCP in mouse, rat, and dog hepatocytes permits HDV infection but does not allow establishment of HBV infection. Contrarily, hepatocytes from cynomolgus macaques, rhesus macaques, and pigs became fully susceptible to HBV upon hNTCP expression with efficiencies comparable to human hepatocytes. Analysis of cloned Ntcp from all species revealed a pronounced role of the human homologue to support HBV and HDV infection. CONCLUSION: Ntcp is the key host factor limiting HBV infection in cynomolgus and rhesus macaques and in pigs. In rodents (mouse, rat) and dogs, transfer of hNTCP supports viral entry but additional host factors are required for the establishment of HBV infection. This finding paves the way for the development of macaques and pigs as immunocompetent animal models to study HBV infection in vivo, immunological responses against the virus and viral pathogenesis. (Hepatology 2017;66:703-716).
Infections with the humanhepatitis B virus (HBV) and hepatitis D virus (HDV) depend on species-specific host factors like the receptor humansodium taurocholate cotransporting polypeptide (hNTCP). Complementation of mouse hepatocytes with hNTCP confers susceptibility to HDV but not HBV, indicating the requirement of additional HBV-specific factors. As an essential premise toward the establishment of an HBV-susceptible animal model, we investigated the role of hNTCP as a limiting factor of hepatocytes in commonly used laboratory animals. Primary hepatocytes from mice, rats, dogs, pigs, rhesus macaques, and cynomolgus macaques were transduced with adeno-associated viral vectors encoding hNTCP and subsequently infected with HBV. Cells were analyzed for Myrcludex B binding, taurocholate uptake, HBV covalently closed circular DNA formation, and expression of all HBV markers. Sodium taurocholate cotransporting polypeptide (Ntcp) from the respective species was cloned and analyzed for HBV and HDV receptor activity in a permissive hepatoma cell line. Expression of hNTCP in mouse, rat, and doghepatocytes permits HDV infection but does not allow establishment of HBV infection. Contrarily, hepatocytes from cynomolgus macaques, rhesus macaques, and pigs became fully susceptible to HBV upon hNTCP expression with efficiencies comparable to human hepatocytes. Analysis of cloned Ntcp from all species revealed a pronounced role of the human homologue to support HBV and HDV infection. CONCLUSION:Ntcp is the key host factor limiting HBV infection in cynomolgus and rhesus macaques and in pigs. In rodents (mouse, rat) and dogs, transfer of hNTCP supports viral entry but additional host factors are required for the establishment of HBV infection. This finding paves the way for the development of macaques and pigs as immunocompetent animal models to study HBV infection in vivo, immunological responses against the virus and viral pathogenesis. (Hepatology 2017;66:703-716).
Authors: Benjamin Y Winer; Elham Shirvani-Dastgerdi; Yaron Bram; Julie Sellau; Benjamin E Low; Heath Johnson; Tiffany Huang; Gabriela Hrebikova; Brigitte Heller; Yael Sharon; Katja Giersch; Sherif Gerges; Kathleen Seneca; Mihai-Alexandru Pais; Angela S Frankel; Luis Chiriboga; John Cullen; Ronald G Nahass; Marc Lutgehetmann; Jared E Toettcher; Michael V Wiles; Robert E Schwartz; Alexander Ploss Journal: Sci Transl Med Date: 2018-06-27 Impact factor: 17.956