Efficient reverse genetics reveals genetic determinants of budding and fusogenic differences between Nipah and Hendra viruses and enables real-time monitoring of viral spread in small animal models of henipavirus infection

Tatyana Yun, Arnold Park, Terence E. Hill, Olivier Pernet, Shannon M. Beaty, Terry L. Juelich, Jennifer K. Smith, Lihong Zhang, Yao E. Wang, Frederic Vigant, Junling Gao, Ping Wu, Benhur Lee, Alexander Freiberg

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Nipah virus (NiV) and Hendra virus (HeV) are closely related henipaviruses of the Paramyxovirinae. Spillover from their fruit bat reservoirs can cause severe disease in humans and livestock. Despite their high sequence similarity, NiV and HeV exhibit apparent differences in receptor and tissue tropism, envelope-mediated fusogenicity, replicative fitness, and other pathophysiologic manifestations. To investigate the molecular basis for these differences, we first established a highly efficient reverse genetics system that increased rescue titers by ≥3 log units, which offset the difficulty of generating multiple recombinants under constraining biosafety level 4 (BSL-4) conditions. We then replaced, singly and in combination, the matrix (M), fusion (F), and attachment glycoprotein (G) genes in mCherry-expressing recombinant NiV (rNiV) with their HeV counterparts. These chimeric but isogenic rNiVs replicated well in primary human endothelial and neuronal cells, indicating efficient heterotypic complementation. The determinants of budding efficiency, fusogenicity, and replicative fitness were dissociable: HeV-M budded more efficiently than NiV-M, accounting for the higher replicative titers of HeV-M-bearing chimeras at early times, while the enhanced fusogenicity of NiV-G-bearing chimeras did not correlate with increased replicative fitness. Furthermore, to facilitate spatiotemporal studies on henipavirus pathogenesis, we generated a firefly luciferase-expressing NiV and monitored virus replication and spread in infected interferon alpha/beta receptor knockout mice via bioluminescence imaging. While intraperitoneal inoculation resulted in neuroinvasion following systemic spread and replication in the respiratory tract, intranasal inoculation resulted in confined spread to regions corresponding to olfactory bulbs and salivary glands before subsequent neuroinvasion. This optimized henipavirus reverse genetics system will facilitate future investigations into the growing numbers of novel henipaviruslike viruses.

Original languageEnglish (US)
Pages (from-to)1242-1253
Number of pages12
JournalJournal of Virology
Volume89
Issue number2
DOIs
StatePublished - 2015

Fingerprint

Henipavirus Infections
Henipavirus
Hendra Virus
Nipah Virus
Nipah virus
Hendra virus
Reverse Genetics
Animal Models
animal models
monitoring
infection
chimerism
Paramyxovirinae
Interferon alpha-beta Receptor
tissue tropism
Firefly Luciferases
livestock diseases
biosafety
receptors
interferon-alpha

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Efficient reverse genetics reveals genetic determinants of budding and fusogenic differences between Nipah and Hendra viruses and enables real-time monitoring of viral spread in small animal models of henipavirus infection. / Yun, Tatyana; Park, Arnold; Hill, Terence E.; Pernet, Olivier; Beaty, Shannon M.; Juelich, Terry L.; Smith, Jennifer K.; Zhang, Lihong; Wang, Yao E.; Vigant, Frederic; Gao, Junling; Wu, Ping; Lee, Benhur; Freiberg, Alexander.

In: Journal of Virology, Vol. 89, No. 2, 2015, p. 1242-1253.

Research output: Contribution to journalArticle

Yun, Tatyana ; Park, Arnold ; Hill, Terence E. ; Pernet, Olivier ; Beaty, Shannon M. ; Juelich, Terry L. ; Smith, Jennifer K. ; Zhang, Lihong ; Wang, Yao E. ; Vigant, Frederic ; Gao, Junling ; Wu, Ping ; Lee, Benhur ; Freiberg, Alexander. / Efficient reverse genetics reveals genetic determinants of budding and fusogenic differences between Nipah and Hendra viruses and enables real-time monitoring of viral spread in small animal models of henipavirus infection. In: Journal of Virology. 2015 ; Vol. 89, No. 2. pp. 1242-1253.
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abstract = "Nipah virus (NiV) and Hendra virus (HeV) are closely related henipaviruses of the Paramyxovirinae. Spillover from their fruit bat reservoirs can cause severe disease in humans and livestock. Despite their high sequence similarity, NiV and HeV exhibit apparent differences in receptor and tissue tropism, envelope-mediated fusogenicity, replicative fitness, and other pathophysiologic manifestations. To investigate the molecular basis for these differences, we first established a highly efficient reverse genetics system that increased rescue titers by ≥3 log units, which offset the difficulty of generating multiple recombinants under constraining biosafety level 4 (BSL-4) conditions. We then replaced, singly and in combination, the matrix (M), fusion (F), and attachment glycoprotein (G) genes in mCherry-expressing recombinant NiV (rNiV) with their HeV counterparts. These chimeric but isogenic rNiVs replicated well in primary human endothelial and neuronal cells, indicating efficient heterotypic complementation. The determinants of budding efficiency, fusogenicity, and replicative fitness were dissociable: HeV-M budded more efficiently than NiV-M, accounting for the higher replicative titers of HeV-M-bearing chimeras at early times, while the enhanced fusogenicity of NiV-G-bearing chimeras did not correlate with increased replicative fitness. Furthermore, to facilitate spatiotemporal studies on henipavirus pathogenesis, we generated a firefly luciferase-expressing NiV and monitored virus replication and spread in infected interferon alpha/beta receptor knockout mice via bioluminescence imaging. While intraperitoneal inoculation resulted in neuroinvasion following systemic spread and replication in the respiratory tract, intranasal inoculation resulted in confined spread to regions corresponding to olfactory bulbs and salivary glands before subsequent neuroinvasion. This optimized henipavirus reverse genetics system will facilitate future investigations into the growing numbers of novel henipaviruslike viruses.",
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AU - Beaty, Shannon M.

AU - Juelich, Terry L.

AU - Smith, Jennifer K.

AU - Zhang, Lihong

AU - Wang, Yao E.

AU - Vigant, Frederic

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AU - Lee, Benhur

AU - Freiberg, Alexander

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