Human glial chimeric mice reveal astrocytic dependence of JC virus infection
Yoichi Kondo, … , Leonid Gorelik, Steven A. Goldman
Yoichi Kondo, … , Leonid Gorelik, Steven A. Goldman
Published November 17, 2014
Citation Information: J Clin Invest. 2014;124(12):5323-5336. https://doi.org/10.1172/JCI76629.
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Research Article

Human glial chimeric mice reveal astrocytic dependence of JC virus infection

Abstract

Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease triggered by infection with the human gliotropic JC virus (JCV). Due to the human-selective nature of the virus, there are no animal models available to investigate JCV pathogenesis. To address this issue, we developed mice with humanized white matter by engrafting human glial progenitor cells (GPCs) into neonatal immunodeficient and myelin-deficient mice. Intracerebral delivery of JCV resulted in infection and subsequent demyelination of these chimeric mice. Human GPCs and astrocytes were infected more readily than oligodendrocytes, and viral replication was noted primarily in human astrocytes and GPCs rather than oligodendrocytes, which instead expressed early viral T antigens and exhibited apoptotic death. Engraftment of human GPCs in normally myelinated and immunodeficient mice resulted in humanized white matter that was chimeric for human astrocytes and GPCs. JCV effectively propagated in these mice, which indicates that astroglial infection is sufficient for JCV spread. Sequencing revealed progressive mutation of the JCV capsid protein VP1 after infection, suggesting that PML may evolve with active infection. These results indicate that the principal CNS targets for JCV infection are astrocytes and GPCs and that infection is associated with progressive mutation, while demyelination is a secondary occurrence, following T antigen–triggered oligodendroglial apoptosis. More broadly, this study provides a model by which to further assess the biology and treatment of human-specific gliotropic viruses.

Authors

Yoichi Kondo, Martha S. Windrem, Lisa Zou, Devin Chandler-Militello, Steven J. Schanz, Romane M. Auvergne, Sarah J. Betstadt, Amy R. Harrington, Mahlon Johnson, Alexander Kazarov, Leonid Gorelik, Steven A. Goldman

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Figure 7

Cell cycle arrest at G2/M in infected glia.

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Cell cycle arrest at G2/M in infected glia. T-Ag+ glia expressed the mit...
T-Ag+ glia expressed the mitosis-associated nuclear protein cyclin B1, as well as the DNA damage and cell cycle arrest–associated phospho-p53(Ser15), regardless of their mitotic stage. (A and B) Vehicle-treated and uninfected (T-Ag–) GPCs expressed intranuclear cyclin B1 only when mitotic and in M phase (the latter as assessed by DAPI; arrows), whereas the nuclei of JCV-infected (T-Ag+) GPCs admitted cyclin B1 in a temporally promiscuous fashion (arrowheads). (C) Nuclear phospho-p53(Ser15) and cyclin B1 were coexpressed by mitotically arrested infected astrocytes, but not by uninfected astrocytes. (D) Like control GPCs, uninfected astrocytes (T-Ag–GFAP+) expressed intranuclear cyclin B1 only in M phase, whereas JCV-infected astrocytes expressed nuclear cyclin B1 even when not dividing. (E and F) JCV-infected astrocytes (T-Ag+GFAP+) coexpressed phospho-p53(Ser15), associated with G/2M arrest, whereas neither vehicle-treated nor T-Ag– astroglia in infected cultures did so to any significant degree. (G and H) Similarly, JCV-infected GPCs (T-Ag+CD140a+) coexpressed phospho-p53(Ser15), as well as did infected oligodendrocytes (T-Ag+O4+) (I and J); for both oligodendrocytes and their progenitors, T-Ag+ cells were significantly more likely to express phospho-p53(Ser15). All cultures were assessed at 10 DPI with type 2A JCV (Mad-1 NCCR). Scale bars: 20 μm. *P < 0.05; ***P < 0.001.