scholarly journals A Virus-Like Particle System Identifies the Endonuclease Domain of Crimean-Congo Hemorrhagic Fever Virus

2015 ◽  
Vol 89 (11) ◽  
pp. 5957-5967 ◽  
Author(s):  
Stephanie Devignot ◽  
Eric Bergeron ◽  
Stuart Nichol ◽  
Ali Mirazimi ◽  
Friedemann Weber
Keyword(s):  
Hemorrhagic Fever ◽  
Fever Virus ◽  
Particle Assembly ◽  
Content Type ◽  
Virus Like Particles ◽  
Crimean Congo Hemorrhagic Fever ◽  
Hemorrhagic Fever Virus ◽  
Virus Like Particle ◽  
Endonuclease Domain ◽  
Biosafety Level

ABSTRACTCrimean-Congo hemorrhagic fever virus(CCHFV; genusNairovirus) is an extremely pathogenic member of theBunyaviridaefamily. 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 predictedin silicoby 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.IMPORTANCECrimean-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.

scholarly journals Assessment of Inhibitors of Pathogenic Crimean-Congo Hemorrhagic Fever Virus Strains Using Virus-Like Particles

2015 ◽  
Vol 9 (12) ◽  
pp. e0004259 ◽  
Author(s):  
Marko Zivcec ◽  
Maureen G. Metcalfe ◽  
César G. Albariño ◽  
Lisa W. Guerrero ◽  
Scott D. Pegan ◽  
...  
Keyword(s):  
Hemorrhagic Fever ◽  
Fever Virus ◽  
Virus Like Particles ◽  
Crimean Congo Hemorrhagic Fever ◽  
Hemorrhagic Fever Virus ◽  
Virus Strains

scholarly journals Similarities in Antiviral Humoral Immune Response to Nucleocapsid Proteins of Hazara and Crimean–Congo Hemorrhagic Fever Virus

Proceedings ◽  
2020 ◽  
Vol 50 (1) ◽  
pp. 72
Author(s):  
Merve Kalkan-Yazıcı ◽  
Mehmet Ziya Doymaz
Keyword(s):  
Humoral Response ◽  
Hemorrhagic Fever ◽  
Cross Reactivity ◽  
Fever Virus ◽  
Nucleocapsid Proteins ◽  
Virus Like Particles ◽  
Crimean Congo Hemorrhagic Fever ◽  
Humoral Immune ◽  
Hemorrhagic Fever Virus ◽  
Transmission Electron

Hazara virus (HAZV), a tick-borne agent of the nairoviruses, is closely related to Crimean–Congo hemorrhagic fever virus (CCHFV). Hazara virus has not been reported as a pathogen for humans and can be studied under BSL-2 conditions, whereas CCHFV causes severe hemorrhagic diseases, with up to 30% mortality rate in humans, and requires BSL-4 facilities to be handled. Serologic and phylogenetic similarities between the two viruses would therefore be an interesting area of research. In this study, we evaluated the immunological similarities between these two viruses using nucleocapsid protein as a model. Here, we evaluated cross-reactivity between CCHFV and HAZV rNP, which forms virus-like particles when expressed in Pichia pastoris. In Western blot assays using CCHFV-infected human and immunized mice and rabbit sera, cross-reactions were detected between the nucleoproteins of both viruses. Virus-like particles were visualized by transmission electron microscopy (TEM) and monitored by dynamic light scattering (DLS). These results suggest that nucleocapsid proteins of HAZV and CCHFV share similarities regarding the antiviral humoral response in both species.

scholarly journals Crimean-Congo Hemorrhagic Fever Virus: Current Advances and Future Prospects of Antiviral Strategies

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1195
Author(s):  
Shiyu Dai ◽  
Fei Deng ◽  
Hualin Wang ◽  
Yunjia Ning
Keyword(s):  
Treatment Options ◽  
Hemorrhagic Fever ◽  
Fever Virus ◽  
Transmission Risk ◽  
High Morbidity ◽  
Tick Bites ◽  
Future Directions ◽  
Crimean Congo Hemorrhagic Fever ◽  
Hemorrhagic Fever Virus ◽  
Biosafety Level

Crimean-Congo hemorrhagic fever virus (CCHFV) is a widespread, tick-borne pathogen that causes Crimean-Congo hemorrhagic fever (CCHF) with high morbidity and mortality. CCHFV is transmitted to humans through tick bites or direct contact with patients or infected animals with viremia. Currently, climate change and globalization have increased the transmission risk of this biosafety level (BSL)-4 virus. The treatment options of CCHFV infection remain limited and there is no FDA-approved vaccine or specific antivirals, which urges the identification of potential therapeutic targets and the design of CCHF therapies with greater effort. In this article, we discuss the current progress and some future directions in the development of antiviral strategies against CCHFV.

scholarly journals Heat Shock Protein 70 Family Members Interact with Crimean-Congo Hemorrhagic Fever Virus and Hazara Virus Nucleocapsid Proteins and Perform a Functional Role in the Nairovirus Replication Cycle

2016 ◽  
Vol 90 (20) ◽  
pp. 9305-9316 ◽  
Author(s):  
Rebecca Surtees ◽  
Stuart D. Dowall ◽  
Amelia Shaw ◽  
Stuart Armstrong ◽  
Roger Hewson ◽  
...  
Keyword(s):  
Life Cycle ◽  
Hemorrhagic Fever ◽  
Therapeutic Strategies ◽  
Fever Virus ◽  
Immunological Methods ◽  
Content Type ◽  
Crimean Congo Hemorrhagic Fever ◽  
Virus Particles ◽  
Hemorrhagic Fever Virus ◽  
Hsp70 Family

ABSTRACTTheNairovirusgenus of theBunyaviridaefamily contains serious human and animal pathogens classified within multiple serogroups and species. Of these serogroups, the Crimean-Congo hemorrhagic fever virus (CCHFV) serogroup comprises sole members CCHFV and Hazara virus (HAZV). CCHFV is an emerging zoonotic virus that causes often-fatal hemorrhagic fever in infected humans for which preventative or therapeutic strategies are not available. In contrast, HAZV is nonpathogenic to humans and thus represents an excellent model to study aspects of CCHFV biology under conditions of more-accessible biological containment. The three RNA segments that form the nairovirus genome are encapsidated by the viral nucleocapsid protein (N) to form ribonucleoprotein (RNP) complexes that are substrates for RNA synthesis and packaging into virus particles. We used quantitative proteomics to identify cellular interaction partners of CCHFV N and identified robust interactions with cellular chaperones. These interactions were validated using immunological methods, and the specific interaction between native CCHFV N and cellular chaperones of the HSP70 family was confirmed during live CCHFV infection. Using infectious HAZV, we showed for the first time that the nairovirus N-HSP70 association was maintained within both infected cells and virus particles, where N is assembled as RNPs. Reduction of active HSP70 levels in cells by the use of small-molecule inhibitors significantly reduced HAZV titers, and a model for chaperone function in the context of high genetic variability is proposed. These results suggest that chaperones of the HSP70 family are required for nairovirus replication and thus represent a genetically stable cellular therapeutic target for preventing nairovirus-mediated disease.IMPORTANCENairoviruses compose a group of human and animal viruses that are transmitted by ticks and associated with serious or fatal disease. One member is Crimean-Congo hemorrhagic fever virus (CCHFV), which is responsible for fatal human disease and is recognized as an emerging threat within Europe in response to climate change. No preventative or therapeutic strategies against nairovirus-mediated disease are currently available. Here we show that the N protein of CCHFV and the related Hazara virus interact with a cellular protein, HSP70, during both the intracellular and extracellular stages of the virus life cycle. The use of inhibitors that block HSP70 function reduces virus titers by up to 1,000-fold, suggesting that this interaction is important within the context of the nairovirus life cycle and may represent a potent target for antinairovirus therapies against which the virus cannot easily develop resistance.

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