Analysis of Hepatitis C Virus Particle Heterogeneity in Immunodeficient Human Liver Chimeric fah-/- Mice

Cell Mol Gastroenterol Hepatol. 2017 Jul 19;4(3):405-417. doi: 10.1016/j.jcmgh.2017.07.002. eCollection 2017 Nov.


Background & aims: Hepatitis C virus (HCV) is a leading cause of chronic liver diseases and the most common indication for liver transplantation in the United States. HCV particles in the blood of infected patients are characterized by heterogeneous buoyant densities, likely owing to HCV association with lipoproteins. However, clinical isolates are not infectious in vitro and the relative infectivity of the particles with respect to their buoyant density therefore cannot be determined, pointing to the need for better in vivo model systems.

Methods: To analyze the evolution of the buoyant density of in vivo-derived infectious HCV particles over time, we infected immunodeficient human liver chimeric fumaryl acetoacetate hydrolase-/- mice with J6/JFH1 and performed ultracentrifugation of infectious mouse sera on isopicnic iodixanol gradients. We also evaluated the impact of a high sucrose diet, which has been shown to increase very-low-density lipoprotein secretion by the liver in rodents, on lipoprotein and HCV particle characteristics.

Results: Similar to the severe combined immunodeficiency disease/Albumin-urokinase plasminogen activator human liver chimeric mouse model, density fractionation of infectious mouse serum showed higher infectivity in the low-density fractions early after infection. However, over the course of the infection, viral particle heterogeneity increased and the overall in vitro infectivity diminished without loss of the human liver graft over time. In mice provided with a sucrose-rich diet we observed a minor shift in HCV infectivity toward lower density that correlated with a redistribution of triglycerides and cholesterol among lipoproteins.

Conclusions: Our work indicates that the heterogeneity in buoyant density of infectious HCV particles evolves over the course of infection and can be influenced by diet.

Keywords: Alb-uPA, Albumin-urokinase plasminogen activator; CETP, cholesterol ester transfer protein; FAH, fumaryl acetoacetate hydrolase; FNRG, absence of fumaryl acetoacetate hydrolase on a immunodeficient NOD Rag gamma IL2 deficient mouse background; FPLC, fast-performance liquid chromatography; HCV; HCV, hepatitis C virus; HCVcc, cell culture–derived hepatitis C virus; HDL, high-density lipoprotein; Human Liver Chimeric Mice; LVP, lipoviroparticle; Lipoprotein; Mouse Model; NRG, nod rag γ; NTBC, nitisinone; PBS, phosphate-buffered saline; SCID, severe combined immunodeficiency disease; VLDL, very low density lipoprotein; apo, apolipoprotein.