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Absence of a robust innate immune response in rat neurons facilitates persistent infection of Borna disease virus in neuronal tissue

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Abstract

Borna disease virus (BDV) persistently infects neurons of the central nervous system of various hosts, including rats. Since type I IFN-mediated antiviral response efficiently blocks BDV replication in primary rat embryo fibroblasts, it has been speculated that BDV is not effectively sensed by the host innate immune system in the nervous system. To test this assumption, organotypical rat hippocampal slice cultures were infected with BDV for up to 4 weeks. This resulted in the secretion of IFN and the up-regulation of IFN-stimulated genes. Using the rat Mx protein as a specific marker for IFN-induced gene expression, astrocytes and microglial cells were found to be Mx positive, whereas neurons, the major cell type in which BDV is replicating, lacked detectable levels of Mx protein. In uninfected cultures, neurons also remained Mx negative even after treatment with high concentrations of IFN-α. This non-responsiveness correlated with a lack of detectable nuclear translocation of both pSTAT1 and pSTAT2 in these cells. Consistently, neuronal dissemination of BDV was not prevented by treatment with IFN-α. These data suggest that the poor innate immune response in rat neurons renders this cell type highly susceptible to BDV infection even in the presence of exogenous IFN-α. Intriguingly, in contrast to rat neurons, IFN-α treatment of mouse neurons resulted in the up-regulation of Mx proteins and block of BDV replication, indicating species-specific differences in the type I IFN response of neurons between mice and rats.

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Acknowledgments

We thank Thomas Michiels, Friedemann Weber, Peter Staeheli, and Georg Kochs for critical reading of the manuscript and Simone Zenker for excellent technical assistance. The study was supported by the Deutsche Forschungsgemeinschaft (DFG) to MS (SCHW 632), BH (He 1520), and in part by the Excellence Initiative of the German Federal and State Governments (GSC-4, Spemann Graduate School).

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Authors and Affiliations

  1. Department of Virology, University of Freiburg, Hermann-Herder-Strasse 11, 79104, Freiburg, Germany

    Chia-Ching Lin, Yuan-Ju Wu & Martin Schwemmle

  2. Spemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, 79104, Freiburg, Germany

    Chia-Ching Lin & Yuan-Ju Wu

  3. Faculty of Biology, University of Freiburg, 79104, Freiburg, Germany

    Chia-Ching Lin & Yuan-Ju Wu

  4. Institute of Anatomy and Cell Biology, University of Freiburg, Albertstrasse 23, 79104, Freiburg, Germany

    Bernd Heimrich

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  1. Chia-Ching Lin
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Correspondence to Bernd Heimrich or Martin Schwemmle.

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18_2013_1402_MOESM1_ESM.tif

Fig. S1 Rat IFN-α/β treatment results in the expression of Mx1 in microglial cells. SD hippocampal slice cultures were either BDV-infected (a) or treated with rat IFN-α/β (103 U/ml) (b). Medium was changed three times per week over 3 weeks. The tissue cultures were fixed on day 21 and stained for the presence of Mx1 (α-Mx1), microglial cells (α-Iba-1) or neurons (α-NeuN). Cell nuclei were visualized with ToPro3. Scale bars, 10 µm (TIFF 4059 kb)

18_2013_1402_MOESM2_ESM.tif

Fig. S2 BDV replication in neurons is not abrogated by treatment with rat type I IFN. (a) BDV-infected hippocampal slice cultures from SD rats were either treated or untreated with rat IFN-α/β (103 U/ml) after 3 days of virus infection. The medium was changed three times per week over 4 weeks. At the indicated time point post-infection, the cell extract was prepared from a pool of at least four cultures and analyzed by Western blotting for the presence of the indicated proteins. (b) Rat hippocampal slice cultures from SD rats were either pretreated with 103 U/ml of rat IFN-α/β for 6 h (IFN pretreated) or left untreated (untreated) and subsequently infected with BDV (1,000 FFU). Medium containing 1,000 U of rat IFN-α/β was changed three times per week in pretreated cultures, whereas no additional IFN was added to the untreated cultures. 21 days post-infection (dpi) viral dissemination was determined by immunofluorescence using BDV-P-specific antibodies. Cell nucleus was visualized by TpPro-3. Scale bars, 10 µm (TIFF 6830 kb)

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Lin, CC., Wu, YJ., Heimrich, B. et al. Absence of a robust innate immune response in rat neurons facilitates persistent infection of Borna disease virus in neuronal tissue. Cell. Mol. Life Sci. 70, 4399–4410 (2013). https://doi.org/10.1007/s00018-013-1402-5

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