Prions hijack tunnelling nanotubes for intercellular spread

  • Nature Cell Biology volume 11, pages 328336 (2009)
  • doi:10.1038/ncb1841
  • Download Citation



In variant Creutzfeldt–Jakob disease, prions (PrPSc) enter the body with contaminated foodstuffs and can spread from the intestinal entry site to the central nervous system (CNS) by intercellular transfer from the lymphoid system to the peripheral nervous system (PNS)1. Although several means2,3,4 and different cell types5,6,7 have been proposed to have a role, the mechanism of cell-to-cell spreading remains elusive. Tunnelling nanotubes (TNTs) have been identified between cells8,9,10,11,12, both in vitro and in vivo10,11,13, and may represent a conserved means of cell-to-cell communication14,15,16. Here we show that TNTs allow transfer of exogenous and endogenous PrPSc between infected and naive neuronal CAD cells17. Significantly, transfer of endogenous PrPSc aggregates was detected exclusively when cells chronically infected with the 139A mouse prion strain were connected to mouse CAD cells by means of TNTs, identifying TNTs as an efficient route for PrPSc spreading in neuronal cells. In addition, we detected the transfer of labelled PrPSc from bone marrow-derived dendritic cells to primary neurons connected through TNTs. Because dendritic cells can interact with peripheral neurons in lymphoid organs, TNT-mediated intercellular transfer would allow neurons to transport prions retrogradely to the CNS1. We therefore propose that TNTs are involved in the spreading of PrPSc within neurons in the CNS and from the peripheral site of entry to the PNS by neuroimmune interactions with dendritic cells.

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  1. 1.

    & Prions and their lethal journey to the brain. Nature Rev. Microbiol. 4, 201–11 (2006).

  2. 2.

    et al. Cells release prions in association with exosomes. Proc. Natl Acad. Sci. USA 101, 9683–9688 (2004).

  3. 3.

    et al. Retrovirus infection strongly enhances scrapie infectivity release in cell culture. EMBO J. 25, 2674–2685 (2006).

  4. 4.

    et al. Intercellular transfer of the cellular prion protein. J. Biol. Chem. 277, 47671–47678 (2002).

  5. 5.

    et al. Infected splenic dendritic cells are sufficient for prion transmission to the CNS in mouse scrapie. J. Clin. Invest. 108, 703–708 (2001).

  6. 6.

    et al. Impaired prion replication in spleens of mice lacking functional follicular dendritic cells. Science 288, 1257–1259 (2000).

  7. 7.

    et al. Lymph nodal prion replication and neuroinvasion in mice devoid of follicular dendritic cells. Proc. Natl Acad. Sci. USA 99, 919–924 (2002).

  8. 8.

    , , , & Nanotubular highways for intercellular organelle transport. Science 303, 1007–1010 (2004).

  9. 9.

    , , & Cutting edge: Membrane nanotubes connect immune cells. J. Immunol. 173, 1511–1513 (2004).

  10. 10.

    , , & Dependence of Drosophila wing imaginal disc cytonemes on Decapentaplegic. Nature 437, 560–563 (2005).

  11. 11.

    & Cytonemes: cellular processes that project to the principal signaling center in Drosophila imaginal discs. Cell 97, 599–607 (1999).

  12. 12.

    et al. Retroviruses can establish filopodial bridges for efficient cell-to-cell transmission. Nature Cell Biol. 9, 310–315 (2007).

  13. 13.

    , & Cutting edge: Membrane nanotubes in vivo: a feature of MHCII+ cells in the mouse cornea. J. Immunol. 180, 5779–5783 (2008).

  14. 14.

    & Apical and lateral cell protrusions interconnect epithelial cells in live Drosophila wing imaginal discs. Dev. Dyn. 236, 3408–3418 (2007).

  15. 15.

    , & Tunneling nanotubes: a new route for the exchange of components between animal cells. FEBS Lett. 581, 2194–2201 (2007).

  16. 16.

    et al. Membrane nanotubes physically connect T cells over long distances presenting a novel route for HIV-1 transmission. Nature Cell Biol. 10, 212–219 (2008).

  17. 17.

    , , & Characterization of a CNS cell line, CAD, in which morphological differentiation is initiated by serum deprivation. J. Neurosci. 17, 1217–1225 (1997).

  18. 18.

    et al. Prion strain discrimination in cell culture: the cell panel assay. Proc. Natl Acad. Sci. USA 104, 20908–20913 (2007).

  19. 19.

    & Cytonemes and tunneling nanotubules in cell–cell communication and viral pathogenesis. Trends Cell Biol. 18, 414–420 (2008).

  20. 20.

    et al. GLUT1CBP(TIP2/GIPC1) interactions with GLUT1 and myosin VI: evidence supporting an adapter function for GLUT1CBP. Mol. Biol. Cell 16, 4183–4201 (2005).

  21. 21.

    Molecular motors: structural adaptations to cellular functions. Nature 389, 561–567 (1997).

  22. 22.

    et al. Uptake and neuritic transport of scrapie prion protein coincident with infection of neuronal cells. J. Neurosci. 25, 5207–5216 (2005).

  23. 23.

    , & Scrapie prion proteins accumulate in the cytoplasm of persistently infected cultured cells. J. Cell Biol. 110, 2117–2132 (1990).

  24. 24.

    & Functional connectivity between immune cells mediated by tunneling nanotubules. Immunity 23, 309–318 (2005).

  25. 25.

    , , , & Migrating intestinal dendritic cells transport PrPSc from the gut. J. Gen. Virol. 83, 267–271 (2002).

  26. 26.

    & Pathological PrP is abundant in sympathetic and sensory ganglia of hamsters fed with scrapie. Neurosci. Lett. 265, 135–138 (1999).

  27. 27.

    Immunologists getting nervous: neuropeptides, dendritic cells and T cell activation. Respir. Res. 2, 133–138 (2001).

  28. 28.

    et al. Interfaces between dendritic cells, other immune cells, and nerve fibres in mouse Peyer's patches: potential sites for neuroinvasion in prion diseases. Microsc. Res. Tech. 66, 1–9 (2005).

  29. 29.

    et al. Oral scrapie infection modifies the homeostasis of Peyer's patches' dendritic cells. Histochem. Cell Biol. 128, 243–251 (2007).

  30. 30.

    et al. Scrapie protein degradation by cysteine proteases in CD11c+ dendritic cells and GT1-1 neuronal cells. J. Virol. 78, 4776–4782 (2004).

  31. 31.

    et al. Packaging of prions into exosomes is associated with a novel pathway of PrP processing. J. Pathol. 211, 582–590 (2007).

  32. 32.

    et al. Transfer of scrapie prion infectivity by cell contact in culture. Curr. Biol. 12, 523–530 (2002).

  33. 33.

    et al. Efficient dissemination of prions through preferential transmission to nearby cells. J. Gen. Virol. 88, 706–713 (2007).

  34. 34.

    et al. Structurally distinct membrane nanotubes between human macrophages support long-distance vesicular traffic or surfing of bacteria. J. Immunol. 177, 8476–8483 (2006).

  35. 35.

    et al. Forced expression of the motor neuron determinant HB9 in neural stem cells affects neurogenesis. Exp. Neurol. 198, 167–182 (2006).

  36. 36.

    , & Prions can infect primary cultured neurons and astrocytes and promote neuronal cell death. Proc. Natl Acad. Sci. USA 101, 12271–12276 (2004).

  37. 37.

    et al. Plasmidic versus insertional cloning of heterologous genes in Mycobacterium bovis BCG: impact on in vivo antigen persistence and immune responses. Infect. Immun. 70, 303–314 (2002).

  38. 38.

    & Culturing hippocampal neurons. Nature Protocols 1, 2406–2415 (2006).

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We thank P. Lazarow, G. Guizzunti and C. Bowler for critical reading of the manuscript. We thank S. Blanchard and D. Bohl-Delfaud for their help in preparing the GFP–PrPwt-retroviral vector, and P. Casanova and J. Vinatier for technical help. We thank H. Laude, A. F. Hill, P. Cossart and M. Way for their gifts (cells, constructs and reagents). We are grateful for assistance with microscopes and image processing received from the Plate-Forme Imagerie Dynamique at the Pasteur Institut. K.G. is supported by the Pasteur Foundation Fellowship Program, E.S. received a fellowship (2004-07) from the Bavarian Research Foundation (BFS), D.B. received funding from the Fondation Canadienne Louis Pasteur, and Z.M. received funding from Ile-de-France. This work was supported by grants to C.Z. from the European Union (Strainbarrier (FP6 Contract No 023183 (Food)) and from Telethon GGP0414.

Author information

Author notes

    • Karine Gousset
    •  & Edwin Schiff

    These authors contributed equally to this work.


  1. Unité de Trafic Membranaire et Pathogénèse, Institut Pasteur, 25 rue du Dr Roux, 75724 Paris Cedex 15, France.

    • Karine Gousset
    • , Edwin Schiff
    • , Christelle Langevin
    • , Zrinka Marijanovic
    • , Anna Caputo
    • , Duncan T. Browman
    •  & Chiara Zurzolo
  2. Department of Immunology, University of Regensburg, F.-J.-Strauss-Allee, 93042 Regensburg, Germany.

    • Edwin Schiff
    •  & Daniela Männel
  3. Dipartimento di Biologia e Patologia Cellulare e Molecolare, Università degli Studi di Napoli 'Federico II', via Pansini 5, 80131 Naples, Italy.

    • Anna Caputo
    •  & Chiara Zurzolo
  4. Unité d'Analyse d'Images Quantitative, Institut Pasteur, 25 rue du Dr Roux, 75724 Paris Cedex 15, France.

    • Nicolas Chenouard
    • , Fabrice de Chaumont
    •  & Jean-Christophe Olivo-Marin
  5. Unité de recherché de Génétique Mycobactérienne, Institut Pasteur, 25 rue du Dr Roux, 75724 Paris Cedex 15, France.

    • Angelo Martino
  6. Groupe “Dynamique des interactions hôte-pathogène”, Institut Pasteur, 25 rue du Dr Roux, 75724 Paris Cedex 15, France.

    • Jost Enninga


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C.Z. and E.S. conceived the project. K.G. and E.S. planned and performed most of the experiments with TNTs in different cells and analysed the data. C.L. planned and performed the infection experiments and analysed the data. Z.M. and A.C. planned and performed experiments in fixed CAD cells and analysed the data. D.T.B. prepared the Alexa-PrPSc and discussed the experiments. N.C. and F.C. performed most of the quantitative image analysis under the supervision of J.C.O. J.E. helped with image reconstruction and discussed the data. A.M. prepared the BMDCs and discussed the related experiments. D.M. co-directed the PhD thesis of E.S. and discussed data with E.S. and C.Z. C.Z. coordinated the project and assisted with planning the experiments and data analysis. K.G., E.S. and C.Z. wrote the manuscript. All authors discussed the results and manuscript text.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Chiara Zurzolo.

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