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Generation of a humanized mouse model with both human immune system and liver cells to model hepatitis C virus infection and liver immunopathogenesis

Nature Protocols volume 7, pages 1608–1617 (2012)Cite this article

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Abstract

Establishing a small animal model that accurately recapitulates hepatotropic pathogens, including hepatitis C virus (HCV) infection and immunopathogenesis, is essential for the study of hepatitis virus–induced liver disease and for therapeutics development. This protocol describes our recently developed humanized mouse model for studying HCV and other hepatotropic infections, human immune response and hepatitis and liver fibrosis. The first 5-h stage is the isolation of human liver progenitor and hematopoietic stem cells from fetal liver. Next, AFC8 immunodeficient mice are transplanted with the isolated progenitor/stem cells. This generally takes 2 h. The transplanted mice are then treated for a month with the mouse liver apoptosis–inducing AFC8 dimerizer and left for an additional 2-month period to permit human liver and immune cell growth as well as system reconstitution and development before inoculation with HCV clinical isolates. HCV infection, human immune response and liver disease are observed with high incidence from approximately 2 months after inoculation.

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Figure 1: AP20187 treatment induces temporal liver–specific damage in AFC8 mice.
Figure 2: Procedure overview: humanization of the liver and immune system in the AFC8 mice.
Figure 3: Human immune system and liver reconstitution in AFC8-hu mice.
Figure 4: Procedure overview: HCV infection and immunopathogenesis.

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Acknowledgements

We thank the Su laboratory members for discussion and support. This work was supported in part by grants from a UNC University Cancer Research Fund innovation grant, from the US National Institutes of Health (AI076142, AA018009 to L.S.) and from a UNC Lineberger Comprehensive Cancer Center Postdoctoral Training grant (M.T.B).

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Authors and Affiliations

  1. Lineberger Comprehensive Cancer Center, University of North Carolina (UNC) at Chapel Hill, Chapel Hill, North Carolina, USA

    Moses T Bility, T Anthony Curtis, Grigoriy I Kovalev & Lishan Su

  2. Center for Infection and Immunity, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China

    Liguo Zhang & Lishan Su

  3. GlaxoSmithKline, Research Triangle Park, North Carolina, USA

    Michael L Washburn

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  1. Moses T Bility
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  2. Liguo Zhang
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  4. T Anthony Curtis
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  5. Grigoriy I Kovalev
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Contributions

M.T.B. and L.S. organized and wrote the article; L.Z., M.L.W., T.A.C. and G.I.K. contributed to writing the article.

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Correspondence to Moses T Bility or Lishan Su.

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Competing interests

The University of North Carolina at Chapel Hill has applied for patents covering parts of the method, with L.Z., M.L.W. and L.S. as coinventors. The University of North Carolina at Chapel Hill manages the invention in accordance with its conflict of interest policies.

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Bility, M., Zhang, L., Washburn, M. et al. Generation of a humanized mouse model with both human immune system and liver cells to model hepatitis C virus infection and liver immunopathogenesis. Nat Protoc 7, 1608–1617 (2012). https://doi.org/10.1038/nprot.2012.083

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