Rescue and characterization of recombinant cedar virus, a non-pathogenic Henipavirus species

Eric D. Laing, Moushimi Amaya, Chanakha K. Navaratnarajah, Yan Ru Feng, Roberto Cattaneo, Lin Fa Wang, Christopher C. Broder

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Background: Hendra virus and Nipah virus are zoonotic viruses that have caused severe to fatal disease in livestock and human populations. The isolation of Cedar virus, a non-pathogenic virus species in the genus Henipavirus, closely-related to the highly pathogenic Hendra virus and Nipah virus offers an opportunity to investigate differences in pathogenesis and receptor tropism among these viruses. Methods: We constructed full-length cDNA clones of Cedar virus from synthetic oligonucleotides and rescued two replication-competent, recombinant Cedar virus variants: a recombinant wild-type Cedar virus and a recombinant Cedar virus that expresses a green fluorescent protein from an open reading frame inserted between the phosphoprotein and matrix genes. Replication kinetics of both viruses and stimulation of the interferon pathway were characterized in vitro. Cellular tropism for ephrin-B type ligands was qualitatively investigated by microscopy and quantitatively by a split-luciferase fusion assay. Results: Successful rescue of recombinant Cedar virus expressing a green fluorescent protein did not significantly affect virus replication compared to the recombinant wild-type Cedar virus. We demonstrated that recombinant Cedar virus stimulated the interferon pathway and utilized the established Hendra virus and Nipah virus receptor, ephrin-B2, but not ephrin-B3 to mediate virus entry. We further characterized virus-mediated membrane fusion kinetics of Cedar virus with the known henipavirus receptors ephrin-B2 and ephrin-B3. Conclusions: The recombinant Cedar virus platform may be utilized to characterize the determinants of pathogenesis across the henipaviruses, investigate their receptor tropisms, and identify novel pan-henipavirus antivirals. Moreover, these experiments can be conducted safely under BSL-2 conditions.

Original languageEnglish (US)
Article number56
JournalVirology Journal
Volume15
Issue number1
DOIs
StatePublished - Mar 27 2018

Fingerprint

Henipavirus
Viruses
Hendra Virus
Nipah Virus
Ephrin-B3
Tropism
Ephrin-B2
Virus Internalization
Green Fluorescent Proteins
Interferons
Ephrins
Virus Receptors

Keywords

  • Cedar virus
  • Ephrin ligands
  • Henipaviruses
  • Paramyxoviridae
  • Receptor tropism
  • Recombinant virus
  • Reverse genetics

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Laing, E. D., Amaya, M., Navaratnarajah, C. K., Feng, Y. R., Cattaneo, R., Wang, L. F., & Broder, C. C. (2018). Rescue and characterization of recombinant cedar virus, a non-pathogenic Henipavirus species. Virology Journal, 15(1), [56]. https://doi.org/10.1186/s12985-018-0964-0

Rescue and characterization of recombinant cedar virus, a non-pathogenic Henipavirus species. / Laing, Eric D.; Amaya, Moushimi; Navaratnarajah, Chanakha K.; Feng, Yan Ru; Cattaneo, Roberto; Wang, Lin Fa; Broder, Christopher C.

In: Virology Journal, Vol. 15, No. 1, 56, 27.03.2018.

Research output: Contribution to journalArticle

Laing, Eric D. ; Amaya, Moushimi ; Navaratnarajah, Chanakha K. ; Feng, Yan Ru ; Cattaneo, Roberto ; Wang, Lin Fa ; Broder, Christopher C. / Rescue and characterization of recombinant cedar virus, a non-pathogenic Henipavirus species. In: Virology Journal. 2018 ; Vol. 15, No. 1.
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abstract = "Background: Hendra virus and Nipah virus are zoonotic viruses that have caused severe to fatal disease in livestock and human populations. The isolation of Cedar virus, a non-pathogenic virus species in the genus Henipavirus, closely-related to the highly pathogenic Hendra virus and Nipah virus offers an opportunity to investigate differences in pathogenesis and receptor tropism among these viruses. Methods: We constructed full-length cDNA clones of Cedar virus from synthetic oligonucleotides and rescued two replication-competent, recombinant Cedar virus variants: a recombinant wild-type Cedar virus and a recombinant Cedar virus that expresses a green fluorescent protein from an open reading frame inserted between the phosphoprotein and matrix genes. Replication kinetics of both viruses and stimulation of the interferon pathway were characterized in vitro. Cellular tropism for ephrin-B type ligands was qualitatively investigated by microscopy and quantitatively by a split-luciferase fusion assay. Results: Successful rescue of recombinant Cedar virus expressing a green fluorescent protein did not significantly affect virus replication compared to the recombinant wild-type Cedar virus. We demonstrated that recombinant Cedar virus stimulated the interferon pathway and utilized the established Hendra virus and Nipah virus receptor, ephrin-B2, but not ephrin-B3 to mediate virus entry. We further characterized virus-mediated membrane fusion kinetics of Cedar virus with the known henipavirus receptors ephrin-B2 and ephrin-B3. Conclusions: The recombinant Cedar virus platform may be utilized to characterize the determinants of pathogenesis across the henipaviruses, investigate their receptor tropisms, and identify novel pan-henipavirus antivirals. Moreover, these experiments can be conducted safely under BSL-2 conditions.",
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