EphrinB2 is the entry receptor for Nipah virus, an emergent deadly paramyxovirus

Article metrics

Abstract

Nipah virus (NiV) is an emergent paramyxovirus that causes fatal encephalitis in up to 70 per cent of infected patients1, and there is evidence of human–to–human transmission2. Endothelial syncytia, comprised of multinucleated giant-endothelial cells, are frequently found in NiV infections, and are mediated by the fusion (F) and attachment (G) envelope glycoproteins. Identification of the receptor for this virus will shed light on the pathobiology of NiV infection, and spur the rational development of effective therapeutics. Here we report that ephrinB2, the membrane-bound ligand for the EphB class of receptor tyrosine kinases (RTKs)3, specifically binds to the attachment (G) glycoprotein of NiV. Soluble Fc-fusion proteins of ephrinB2, but not ephrinB1, effectively block NiV fusion and entry into permissive cell types. Moreover, transfection of ephrinB2 into non-permissive cells renders them permissive for NiV fusion and entry. EphrinB2 is expressed on endothelial cells and neurons3,4, which is consistent with the known cellular tropism for NiV5. Significantly, we find that NiV-envelope-mediated infection of microvascular endothelial cells and primary cortical rat neurons is inhibited by soluble ephrinB2, but not by the related ephrinB1 protein. Cumulatively, our data show that ephrinB2 is a functional receptor for NiV.

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: Soluble NiV-G binds to a 48 kDa membrane protein.
Figure 2: The ectodomain of NiV-G binds specifically to ephrinB2.
Figure 3: EphrinB2 is necessary for NiV fusion.
Figure 4: EphrinB2 mediates entry of NiV-F/G pseudotyped viruses.

References

  1. 1

    Hsu, V. P. et al. Nipah virus encephalitis reemergence, Bangladesh. Emerg. Infect. Dis. 10, 2082–2087 (2004)

  2. 2

    The International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDRB). Person-to-person transmission of Nipah virus during outbreak in Faridpur District. Health Sci. Bull. 2, 5–9 (2004)

  3. 3

    Poliakov, A., Cotrina, M. & Wilkinson, D. G. Diverse roles of eph receptors and ephrins in the regulation of cell migration and tissue assembly. Dev. Cell 7, 465–480 (2004)

  4. 4

    Palmer, A. & Klein, R. Multiple roles of ephrins in morphogenesis, neuronal networking, and brain function. Genes Dev. 17, 1429–1450 (2003)

  5. 5

    Wong, K. T. et al. Nipah virus infection: pathology and pathogenesis of an emerging paramyxoviral zoonosis. Am. J. Pathol. 161, 2153–2167 (2002)

  6. 6

    Chua, K. B. et al. Nipah virus: a recently emergent deadly paramyxovirus. Science 288, 1432–1435 (2000)

  7. 7

    Harcourt, B. H. et al. Molecular characterization of Nipah virus, a newly emergent paramyxovirus. Virology 271, 334–349 (2000)

  8. 8

    Parashar, U. D. et al. Case-control study of risk factors for human infection with a new zoonotic paramyxovirus, Nipah virus, during a 1998–1999 outbreak of severe encephalitis in Malaysia. J. Infect. Dis. 181, 1755–1759 (2000)

  9. 9

    Lam, S. K. Nipah virus–a potential agent of bioterrorism? Antiviral Res. 57, 113–119 (2003)

  10. 10

    Field, H. et al. The natural history of Hendra and Nipah viruses. Microbes Infect. 3, 307–314 (2001)

  11. 11

    Wong, K. T. et al. A golden hamster model for human acute Nipah virus infection. Am. J. Pathol. 163, 2127–2137 (2003)

  12. 12

    Hooper, P., Zaki, S., Daniels, P. & Middleton, D. Comparative pathology of the diseases caused by Hendra and Nipah viruses. Microbes Infect. 3, 315–322 (2001)

  13. 13

    Bossart, K. N. et al. Membrane fusion tropism and heterotypic functional activities of the Nipah virus and Hendra virus envelope glycoproteins. J. Virol. 76, 11186–11198 (2002)

  14. 14

    Tamin, A. et al. Functional properties of the fusion and attachment glycoproteins of Nipah virus. Virology 296, 190–200 (2002)

  15. 15

    Guillaume, V. et al. Nipah virus: vaccination and passive protection studies in a hamster model. J. Virol. 78, 834–840 (2004)

  16. 16

    Gale, N. W. et al. Ephrin-B2 selectively marks arterial vessels and neovascularization sites in the adult, with expression in both endothelial and smooth-muscle cells. Dev. Biol. 230, 151–160 (2001)

  17. 17

    Shin, D. et al. Expression of ephrinB2 identifies a stable genetic difference between arterial and venous vascular smooth muscle as well as endothelial cells, and marks subsets of microvessels at sites of adult neovascularization. Dev. Biol. 230, 139–150 (2001)

  18. 18

    Rucker, J. et al. Cell-cell fusion assay to study role of chemokine receptors in human immunodeficiency virus type 1 entry. Methods Enzymol. 288, 118–133 (1997)

  19. 19

    Bossart, K. N. & Broder, C. C. Viral glycoprotein-mediated cell fusion assays using vaccinia virus vectors. Methods Mol. Biol. 269, 309–332 (2004)

  20. 20

    Takada, A. et al. A system for functional analysis of Ebola virus glycoprotein. Proc. Natl Acad. Sci. USA 94, 14764–14769 (1997)

  21. 21

    Esko, J. D., Stewart, T. E. & Taylor, W. H. Animal cell mutants defective in glycosaminoglycan biosynthesis. Proc. Natl Acad. Sci. USA 82, 3197–3201 (1985)

  22. 22

    Liu, J. & Thorp, S. C. Cell surface heparan sulfate and its roles in assisting viral infections. Med. Res. Rev. 22, 1–25 (2002)

  23. 23

    Kullander, K. & Klein, R. Mechanisms and functions of Eph and ephrin signalling. Nature Rev. Mol. Cell Biol. 3, 475–486 (2002)

  24. 24

    Noren, N. K., Lu, M., Freeman, A. L., Koolpe, M. & Pasquale, E. B. Interplay between EphB4 on tumour cells and vascular ephrin-B2 regulates tumour growth. Proc. Natl Acad. Sci. USA 101, 5583–5588 (2004)

  25. 25

    Estus, S. et al. Aggregated amyloid-beta protein induces cortical neuronal apoptosis and concomitant “apoptotic” pattern of gene induction. J. Neurosci. 17, 7736–7745 (1997)

  26. 26

    Shao, R. & Guo, X. Human microvascular endothelial cells immortalized with human telomerase catalytic protein: a model for the study of in vitro angiogenesis. Biochem. Biophys. Res. Commun. 321, 788–794 (2004)

  27. 27

    Tollefsen, S. et al. DNA injection in combination with electroporation: a novel method for vaccination of farmed ruminants. Scand. J. Immunol. 57, 229–238 (2003)

Download references

Acknowledgements

We thank M. Whitt for permission to use the VSV-pseudotype system, B. Reversade for discussions, and K. Adams for editorial comments. This work was supported by NIH grants to B.L., an NIH NRSA grant to O.A.N., an emerging infectious disease training grant to A.B.-C., and a biodefence research fellowship to E.L.L. S.T. was supported by an NSF-funded UCLA-IGERT bioinformatics traineeship. B.L. is a recipient of the Burroughs Wellcome Fund Career Development Award and is also a Charles E. Culpepper Medical Scholar supported by the Rockefeller Brothers Fund and by Goldman Philanthropic Partnerships. We also acknowledge support from the UCLA AIDS Institute and the flow cytometry core (UCLA CFAR). Author Contributions E.L.L., H.C.A., A.B.-C. and R.N. contributed equally to this work.

Author information

Correspondence to Benhur Lee.

Ethics declarations

Competing interests

Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Supplementary information

Supplementary Figure 1

This figure contains the protein sequence of EphrinB2; highlighting the two peptide fragments that were identified by LC-MS/MS (tandem MS). (DOC 19 kb)

Supplementary Table 1

Table I contains a list of deletions made in NiV-G that lead to the identification of δ28 NiV-G-Fc. (DOC 31 kb)

Supplementary Methods 1

The methods used for the construction, production and measurement of NiV-G-Fc and sNiV-G-HA are outlined here. (DOC 25 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Negrete, O., Levroney, E., Aguilar, H. et al. EphrinB2 is the entry receptor for Nipah virus, an emergent deadly paramyxovirus. Nature 436, 401–405 (2005) doi:10.1038/nature03838

Download citation

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.