Mice Expressing Minimally Humanized CD81 and Occludin Genes Support Hepatitis C Virus Uptake In Vivo.

TitleMice Expressing Minimally Humanized CD81 and Occludin Genes Support Hepatitis C Virus Uptake In Vivo.
Publication TypeJournal Article
Year of Publication2017
AuthorsDing, Q, von Schaewen, M, Hrebikova, G, Heller, B, Sandmann, L, Plaas, M, Ploss, A
JournalJ Virol
Volume91
Issue4
Date Published2017 02 15
ISSN1098-5514
KeywordsAlleles, Amino Acid Sequence, Animals, Disease Models, Animal, Gene Expression, Gene Targeting, Genes, Reporter, Genetic Loci, Hepacivirus, Hepatitis C, Host-Pathogen Interactions, Humans, Mice, Mice, Transgenic, Occludin, Organ Specificity, Phenotype, Tetraspanin 28, Transcription, Genetic, Virus Internalization
Abstract

<p>Hepatitis C virus (HCV) causes chronic infections in at least 150 million individuals worldwide. HCV has a narrow host range and robustly infects only humans and chimpanzees. The underlying mechanisms for this narrow host range are incompletely understood. At the level of entry, differences in the amino acid sequences between the human and mouse orthologues of two essential host factors, the tetraspanin CD81 and the tight junction protein occludin (OCLN), explain, at least in part, HCV's limited ability to enter mouse hepatocytes. We have previously shown that adenoviral or transgenic overexpression of human CD81 and OCLN facilitates HCV uptake into mouse hepatocytes in vitro and in vivo In efforts to refine these models, we constructed knock-in mice in which the second extracellular loops of CD81 and OCLN were replaced with the respective human sequences, which contain the determinants that are critical for HCV uptake. We demonstrate that the humanized CD81 and OCLN were expressed at physiological levels in a tissue-appropriate fashion. Mice bearing the humanized alleles formed normal tight junctions and did not exhibit any immunologic abnormalities, indicating that interactions with their physiological ligands were intact. HCV entry factor knock-in mice take up HCV with an efficiency similar to that in mice expressing HCV entry factors transgenically or adenovirally, demonstrating the utility of this model for studying HCV infection in vivo IMPORTANCE: At least 150 million individuals are chronically infected with hepatitis C virus (HCV). Chronic hepatitis C can result in progressive liver disease and liver cancer. New antiviral treatments can cure HCV in the majority of patients, but a vaccine remains elusive. To gain a better understanding of the processes culminating in liver failure and cancer and to prioritize vaccine candidates more efficiently, small-animal models are needed. Here, we describe the characterization of a new mouse model in which the parts of two host factors that are essential for HCV uptake, CD81 and occludin (OCLN), which differ between mice and humans, were humanized. We demonstrate that such minimally humanized mice develop normally, express the modified genes at physiological levels, and support HCV uptake. This model is of considerable utility for studying viral entry in the three-dimensional context of the liver and to test approaches aimed at preventing HCV entry.</p>

DOI10.1128/JVI.01799-16
Alternate JournalJ Virol
PubMed ID27928007
PubMed Central IDPMC5286898
Grant ListR01 AI079031 / AI / NIAID NIH HHS / United States
R01 AI107301 / AI / NIAID NIH HHS / United States
R21 AI117213 / AI / NIAID NIH HHS / United States