Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Prions hijack tunnelling nanotubes for intercellular spread

Abstract

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.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: TNT analyses in mouse neuronal CAD cells.
Figure 2: LysoTracker-labelled vesicles and GFP–PrPwt transfer through TNTs between CAD cells.
Figure 3: Brain homogenate and infectious Alexa-PrPSc transfer through TNTs.
Figure 4: Detection and quantification of endogenous PrPSc transfer in CAD and ScCAD cells through TNTs.
Figure 5: BMDCs can interact with primary neurons through TNTs to spread infection.

References

  1. 1

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

    CAS  Article  Google Scholar 

  2. 2

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

    CAS  Article  Google Scholar 

  3. 3

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

    CAS  Article  Google Scholar 

  4. 4

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

    CAS  Article  Google Scholar 

  5. 5

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

    CAS  Article  Google Scholar 

  6. 6

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

    CAS  Article  Google Scholar 

  7. 7

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

    CAS  Article  Google Scholar 

  8. 8

    Rustom, A., Saffrich, R., Markovic, I., Walther, P. & Gerdes, H. H. Nanotubular highways for intercellular organelle transport. Science 303, 1007–1010 (2004).

    CAS  Article  Google Scholar 

  9. 9

    Onfelt, B., Nedvetzki, S., Yanagi, K. & Davis, D. M. Cutting edge: Membrane nanotubes connect immune cells. J. Immunol. 173, 1511–1513 (2004).

    Article  Google Scholar 

  10. 10

    Hsiung, F., Ramirez-Weber, F. A., Iwaki, D. D. & Kornberg, T. B. Dependence of Drosophila wing imaginal disc cytonemes on Decapentaplegic. Nature 437, 560–563 (2005).

    CAS  Article  Google Scholar 

  11. 11

    Ramirez-Weber, F. A. & Kornberg, T. B. Cytonemes: cellular processes that project to the principal signaling center in Drosophila imaginal discs. Cell 97, 599–607 (1999).

    CAS  Article  Google Scholar 

  12. 12

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

    CAS  Article  Google Scholar 

  13. 13

    Chinnery, H. R., Pearlman, E. & McMenamin, P. G. Cutting edge: Membrane nanotubes in vivo: a feature of MHCII+ cells in the mouse cornea. J. Immunol. 180, 5779–5783 (2008).

    CAS  Article  Google Scholar 

  14. 14

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

    CAS  Article  Google Scholar 

  15. 15

    Gerdes, H.-H., Bukoreshtliev, N. V. & Barroso, J. F. V. Tunneling nanotubes: a new route for the exchange of components between animal cells. FEBS Lett. 581, 2194–2201 (2007).

    CAS  Article  Google Scholar 

  16. 16

    Sowinski, S. 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).

    Article  Google Scholar 

  17. 17

    Qi, Y., Wang, J. K. T., McMillian, M. & Chikaraishi, D. M. Characterization of a CNS cell line, CAD, in which morphological differentiation is initiated by serum deprivation. J. Neurosci. 17, 1217–1225 (1997).

    CAS  Article  Google Scholar 

  18. 18

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

    CAS  Article  Google Scholar 

  19. 19

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

    CAS  Article  Google Scholar 

  20. 20

    Reed, B. C. 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).

    CAS  Article  Google Scholar 

  21. 21

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

    CAS  Article  Google Scholar 

  22. 22

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

    CAS  Article  Google Scholar 

  23. 23

    Taraboulos, A., Serban, D. & Prusiner, S. B. Scrapie prion proteins accumulate in the cytoplasm of persistently infected cultured cells. J. Cell Biol. 110, 2117–2132 (1990).

    CAS  Article  Google Scholar 

  24. 24

    Watkins, S. C. & Salter, R. D. Functional connectivity between immune cells mediated by tunneling nanotubules. Immunity 23, 309–318 (2005).

    CAS  Article  Google Scholar 

  25. 25

    Huang, F. P., Farquhar, C. F., Mabbott, N. A., Bruce, M. E. & MacPherson, G. G. Migrating intestinal dendritic cells transport PrPSc from the gut. J. Gen. Virol. 83, 267–271 (2002).

    CAS  Article  Google Scholar 

  26. 26

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

    CAS  Article  Google Scholar 

  27. 27

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

    CAS  Article  Google Scholar 

  28. 28

    Defaweux, V. 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).

    CAS  Article  Google Scholar 

  29. 29

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

    CAS  Article  Google Scholar 

  30. 30

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

    CAS  Article  Google Scholar 

  31. 31

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

    CAS  Article  Google Scholar 

  32. 32

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

    CAS  Article  Google Scholar 

  33. 33

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

    CAS  Article  Google Scholar 

  34. 34

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

    Article  Google Scholar 

  35. 35

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

    CAS  Article  Google Scholar 

  36. 36

    Cronier, S., Laude, H. & Peyrin, J. M. Prions can infect primary cultured neurons and astrocytes and promote neuronal cell death. Proc. Natl Acad. Sci. USA 101, 12271–12276 (2004).

    CAS  Article  Google Scholar 

  37. 37

    Mederle, I. 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).

    CAS  Article  Google Scholar 

  38. 38

    Kaech, S. & Banker, G. Culturing hippocampal neurons. Nature Protocols 1, 2406–2415 (2006).

    CAS  Article  Google Scholar 

Download references

Acknowledgements

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

Affiliations

Authors

Contributions

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.

Corresponding author

Correspondence to Chiara Zurzolo.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary Information (PDF 1520 kb)

Supplementary Information

Supplementary Movie 1 (MOV 3247 kb)

Supplementary Information

Supplementary Movie 2 (MOV 3390 kb)

Supplementary Information

Supplementary Movie 3 (MOV 5187 kb)

Supplementary Information

Supplementary Movie 4 (MOV 108 kb)

Supplementary Information

Supplementary Movie 5 (MOV 755 kb)

Supplementary Information

Supplementary Movie 6 (MOV 750 kb)

Supplementary Information

Supplementary Movie 7 (MOV 5175 kb)

Supplementary Information

Supplementary Movie 8 (MOV 518 kb)

Supplementary Information

Supplementary Movie 9 (MOV 4558 kb)

Supplementary Information

Supplementary Movie 10 (MOV 5476 kb)

Supplementary Information

Supplementary Movie 11 (MOV 4329 kb)

Supplementary Information

Supplementary Movie 12 (MOV 724 kb)

Supplementary Information

Supplementary Movie 13 (MOV 755 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Gousset, K., Schiff, E., Langevin, C. et al. Prions hijack tunnelling nanotubes for intercellular spread. Nat Cell Biol 11, 328–336 (2009). https://doi.org/10.1038/ncb1841

Download citation

Further reading

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing