Rescue and characterization of recombinant cedar virus, a non-pathogenic Henipavirus species
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RESEARCH
Open Access
Rescue and characterization of recombinant cedar virus, a non-pathogenic Henipavirus species Eric D. Laing1†, Moushimi Amaya1†, Chanakha K. Navaratnarajah2, Yan-Ru Feng1, Roberto Cattaneo2, Lin-Fa Wang3 and Christopher C. Broder1*
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, closelyrelated 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. Keywords: Cedar virus, Henipaviruses, Paramyxoviridae, Ephrin ligands, Reverse genetics, Recombinant virus, Receptor tropism
Background Hendra virus (HeV) and Nipah virus (NiV) are the prototypical viruses of the genus Henipavirus [1], and are notable highly pathogenic zoonotic paramyxoviruses that have caused numerous severe and often fatal acute respiratory and/or neurologic disease in humans and livestock since their initial recognition in Australia * Correspondence: [email protected] † Equal contributors 1 Department of Microbiology and Immunology, Uniformed Services University, Bethesda, MD 20814, USA Full list of author information is available at the end of the article
(HeV) and Malaysia, Bangladesh, India and the Philippines (NiV) (reviewed in: [2–4]). The identification of HeV following fatal respiratory illness in 17 horses and one human in 1994 [5], was followed by the first recorded spillovers of NiV between 1998 and 1999
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