Abstract:
UNLABELLED:Crimean-Congo hemorrhagic fever virus (CCHFV; genus Nairovirus) is an extremely pathogenic member of the Bunyaviridae family. Since handling of the virus requires a biosafety level 4 (BSL-4) facility, little is known about pathomechanisms and host interactions. Here, we describe the establishment of a transcriptionally competent virus-like particle (tc-VLP) system for CCHFV. Recombinant polymerase (L), nucleocapsid protein (N) and a reporter minigenome expressed in human HuH-7 cells resulted in formation of transcriptionally active nucleocapsids that could be packaged by coexpressed CCHFV glycoproteins into tc-VLPs. The tc-VLPs resembled authentic virus particles in their protein composition and neutralization sensitivity to anti-CCHFV antibodies and could recapitulate all steps of the viral replication cycle. Particle attachment, entry, and primary transcription were modeled by infection of naive cells. The subsequent steps of genome replication, secondary transcription, and particle assembly and release can be obtained upon passaging the tc-VLPs on cells expressing CCHFV structural proteins. The utility of the VLP system was demonstrated by showing that the endonuclease domain of L is located around amino acid D693, as was predicted in silico by B. Morin et al. (PLoS Pathog 6:e1001038, 2010, http://dx.doi.org/10.1371/journal.ppat.1001038). The tc-VLP system will greatly facilitate studies and diagnostics of CCHFV under non-BSL-4 conditions. IMPORTANCE:Crimean-Congo hemorrhagic fever virus (CCHFV) is an extremely virulent pathogen of humans. Since the virus can be handled only at the highest biosafety level, research is restricted to a few specialized laboratories. We developed a plasmid-based system to produce virus-like particles with the ability to infect cells and transcribe a reporter genome. Due to the absence of viral genes, the virus-like particles are unable to spread or cause disease, thus allowing study of aspects of CCHFV biology under relaxed biosafety conditions.
journal_name
J Viroljournal_title
Journal of virologyauthors
Devignot S,Bergeron E,Nichol S,Mirazimi A,Weber Fdoi
10.1128/JVI.03691-14subject
Has Abstractpub_date
2015-06-01 00:00:00pages
5957-67issue
11eissn
0022-538Xissn
1098-5514pii
JVI.03691-14journal_volume
89pub_type
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pub_type: 杂志文章
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更新日期:1970-03-01 00:00:00
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pub_type: 杂志文章
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更新日期:1998-02-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
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更新日期:1977-03-01 00:00:00
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pub_type: 临床试验,杂志文章
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更新日期:1994-12-01 00:00:00
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pub_type: 杂志文章
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pub_type: 杂志文章
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journal_title:Journal of virology
pub_type: 杂志文章
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journal_title:Journal of virology
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更新日期:1990-04-01 00:00:00
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pub_type: 杂志文章
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更新日期:1978-01-01 00:00:00
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pub_type: 杂志文章
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pub_type: 杂志文章
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