Antagonistic Antiviral Activity between IFN-Lambda and IFN-Alpha against Lethal Crimean-Congo Hemorrhagic Fever Virus In Vitro

AbstractCrimean-Congo hemorrhagic fever virus (CCHFV) is one of the prioritized diseases of World Health Organization, considering its potential to create a public health emergency and more importantly, the absence of efficacious drugs and/or vaccines regarding treatment. The highly lethal nature characteristic to CCHFV restricts research to BSL-4 laboratories, which complicates effective research and developmental strategies. In consideration of antiviral therapies, RNA interference can be used to suppress viral replication by targeting viral genes. RNA interference uses small interfering RNAs (siRNAs) to silence genes. The aim of our study was to design siRNAs that inhibit CCHFV replication and can serve as a basis for further antiviral therapies. A549 cells were infected with CCHFV after transfection with the siRNAs. Following 72 hours, nucleic acid from the supernatant was extracted for Droplet Digital PCR analysis. Among the investigated siRNAs we identified four effective candidates against all three segments of CCHF genome: one for the S and M segments, whilst two for the L segment. Consequently, blocking any segment of CCHFV leads to changes in the virus copy number that indicates an antiviral effect of the siRNAs in vitro. The most active siRNAs were demonstrated a specific inhibitory effect against CCHFV in a dose-dependent manner. In summary, we demonstrated the ability of specific siRNAs to inhibit CCHFV replication in vitro. This promising result can be used in future anti-CCHFV therapy developments.

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Small Interfering RNAs Are Highly Effective Inhibitors of Crimean-Congo Hemorrhagic Fever Virus Replication In Vitro

Molecules ◽

10.3390/molecules25235771 ◽

2020 ◽

Vol 25 (23) ◽

pp. 5771

Author(s):

Fanni Földes ◽

Mónika Madai ◽

Henrietta Papp ◽

Gábor Kemenesi ◽

Brigitta Zana ◽

...

Keyword(s):

Rna Interference ◽

A549 Cells ◽

Hemorrhagic Fever ◽

Fever Virus ◽

World Health ◽

Small Interfering Rnas ◽

Crimean Congo Hemorrhagic Fever ◽

Hemorrhagic Fever Virus ◽

Antiviral Therapies

Crimean-Congo hemorrhagic fever virus (CCHFV) is one of the prioritized diseases of the World Health Organization, considering its potential to create a public health emergency and, more importantly, the absence of efficacious drugs and/or vaccines for treatment. The highly pathogenic characteristic of CCHFV restricts research to BSL-4 laboratories, which complicates effective research and developmental strategies. In consideration of antiviral therapies, RNA interference can be used to suppress viral replication by targeting viral genes. RNA interference uses small interfering RNAs (siRNAs) to silence genes. The aim of our study was to design and test siRNAs in vitro that inhibit CCHFV replication and can serve as a basis for further antiviral therapies. A549 cells were infected with CCHFV after transfection with the siRNAs. Following 72 h, nucleic acid from the supernatant was extracted for RT Droplet Digital PCR analysis. Among the investigated siRNAs we identified effective candidates against all three segments of the CCHF genome. Consequently, blocking any segment of CCHFV leads to changes in the virus copy number that indicates an antiviral effect of the siRNAs. In summary, we demonstrated the ability of specific siRNAs to inhibit CCHFV replication in vitro. This promising result can be integrated into future anti-CCHFV therapy developments.

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Evaluation of Crimean-Congo hemorrhagic fever virus in vitro inhibition by chloroquine and chlorpromazine, two FDA approved molecules

Antiviral Research ◽

10.1016/j.antiviral.2015.03.005 ◽

2015 ◽

Vol 118 ◽

pp. 75-81 ◽

Cited By ~ 25

Author(s):

O. Ferraris ◽

M. Moroso ◽

O. Pernet ◽

S. Emonet ◽

A. Ferrier Rembert ◽

...

Keyword(s):

Hemorrhagic Fever ◽

Fever Virus ◽

Crimean Congo Hemorrhagic Fever ◽

Hemorrhagic Fever Virus ◽

In Vitro Inhibition

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The Crimean-Congo Hemorrhagic Fever Virus NSm Protein Is Dispensable for Growth In Vitro and Disease in Ifnar-/- Mice

Microorganisms ◽

10.3390/microorganisms8050775 ◽

2020 ◽

Vol 8 (5) ◽

pp. 775 ◽

Cited By ~ 1

Author(s):

Stephen R. Welch ◽

Florine E. M. Scholte ◽

Jessica R. Spengler ◽

Jana M. Ritter ◽

JoAnn D. Coleman-McCray ◽

...

Keyword(s):

Clinical Signs ◽

Hemorrhagic Fever ◽

Fever Virus ◽

Structural Proteins ◽

Glycoprotein Precursor ◽

Crimean Congo Hemorrhagic Fever ◽

Virus Attachment ◽

Hemorrhagic Fever Virus ◽

Endoproteolytic Cleavage

Crimean-Congo hemorrhagic fever virus (CCHFV) is a tri-segmented, tick-borne nairovirus that causes disease of ranging severity in humans. The CCHFV M segment encodes a complex glycoprotein precursor (GPC) that undergoes extensive endoproteolytic cleavage, giving rise to two structural proteins (Gn and Gc) required for virus attachment and entry, and to multiple non-structural proteins (NSm, GP160, GP85, and GP38). The functions of these non-structural proteins remain largely unclear. Here, we investigate the role of NSm during infection by generating a recombinant CCHFV lacking the complete NSm domain (10200∆NSm) and observing CCHFV ∆NSm replication in cell lines and pathogenicity in Ifnar-/- mice. Our data demonstrate that the NSm domain is dispensable for viral replication in vitro, and, despite the delayed onset of clinical signs, CCHFV lacking this domain caused severe or lethal disease in infected mice.

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Comparison of Antiviral Activity of Recombinant and Natural Interferons Against Crimean-Congo Hemorrhagic Fever Virus~!2009-11-09~!2009-12-03~!2010-04-22~!

The Open Virology Journal ◽

10.2174/1874357901004020038 ◽

2010 ◽

Vol 4 (2) ◽

pp. 38-41 ◽

Cited By ~ 1

Author(s):

Helen Karlberg ◽

Gunnel Lindegren ◽

Ali Mirazimi

Keyword(s):

Antiviral Activity ◽

Hemorrhagic Fever ◽

Fever Virus ◽

Crimean Congo Hemorrhagic Fever ◽

Hemorrhagic Fever Virus

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Human MxA Protein Inhibits the Replication of Crimean-Congo Hemorrhagic Fever Virus

Journal of Virology ◽

10.1128/jvi.78.8.4323-4329.2004 ◽

2004 ◽

Vol 78 (8) ◽

pp. 4323-4329 ◽

Cited By ~ 78

Author(s):

Ida Andersson ◽

Linda Bladh ◽

Mehrdad Mousavi-Jazi ◽

Karl-Eric Magnusson ◽

Åke Lundkvist ◽

...

Keyword(s):

Antiviral Activity ◽

Hemorrhagic Fever ◽

Inhibitory Effect ◽

Fever Virus ◽

Progeny Virus ◽

Crimean Congo Hemorrhagic Fever ◽

Viral Genomes ◽

Virus Particles ◽

Hemorrhagic Fever Virus ◽

Infected Cells

ABSTRACT Crimean-Congo hemorrhagic fever virus (CCHFV) belongs to the genus Nairovirus within the family Bunyaviridae and is the causative agent of severe hemorrhagic fever. Despite increasing knowledge about hemorrhagic fever viruses, the factors determining their pathogenicity are still poorly understood. The interferon-induced MxA protein has been shown to have an inhibitory effect on several members of the Bunyaviridae family, but the effect of MxA against CCHFV has not previously been studied. Here, we report that human MxA has antiviral activity against CCHFV. The yield of progeny virus in cells constitutively expressing MxA was reduced up to 1,000-fold compared with control cells, and accumulation of viral genomes was blocked. Confocal microscopy revealed that MxA colocalizes with the nucleocapsid protein (NP) of CCHFV in the perinuclear regions of infected cells. Furthermore, we found that MxA interacted with NP by using a coimmunoprecipitation assay. We also found that an amino acid substitution (E645R) within the C-terminal domain of MxA resulted in a loss of MxA antiviral activity and, concomitantly, in the capacity to interact with CCHFV NP. These results suggest that MxA, by interacting with a component of the nucleocapsid, prevents replication of CCHFV viral RNA and thereby inhibits the production of new infectious virus particles.

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Differential Growth Characteristics of Crimean-Congo Hemorrhagic Fever Virus in Kidney Cells of Human and Bovine Origin

Viruses ◽

10.3390/v12060685 ◽

2020 ◽

Vol 12 (6) ◽

pp. 685

Author(s):

Katalin Földes ◽

Touraj Aligholipour Farzani ◽

Koray Ergünay ◽

Aykut Ozkul

Keyword(s):

Cell Lines ◽

Human Kidney ◽

Hemorrhagic Fever ◽

Fever Virus ◽

Kidney Cells ◽

Bovine Kidney ◽

Crimean Congo Hemorrhagic Fever ◽

Hemorrhagic Fever Virus ◽

The Impact

Crimean-Congo hemorrhagic fever virus (CCHFV) causes a lethal tick-borne zoonotic disease with severe clinical manifestation in humans but does not produce symptomatic disease in wild or domestic animals. The factors contributing to differential outcomes of infection between species are not yet understood. Since CCHFV is known to have tropism to kidney tissue and cattle play an important role as an amplifying host for CCHFV, in this study, we assessed in vitro cell susceptibility to CCHFV infection in immortalized and primary kidney and adrenal gland cell lines of human and bovine origin. Based on our indirect fluorescent focus assay (IFFA), we suggest a cell-to-cell CCHF viral spread process in bovine kidney cells but not in human cells. Over the course of seven days post-infection (dpi), infected bovine kidney cells are found in restricted islet-like areas. In contrast, three dpi infected human kidney or adrenal cells were noted in areas distant from one another yet progressed to up to 100% infection of the monolayer. Pronounced CCHFV replication, measured by quantitative real-time RT-PCR (qRT-PCR) of both intra- and extracellular viral RNA, was documented only in human kidney cells, supporting restrictive infection in cells of bovine origin. To further investigate the differences, lactate dehydrogenase activity and cytopathic effects were measured at different time points in all mentioned cells. In vitro assays indicated that CCHFV infection affects human and bovine kidney cells differently, where human cell lines seem to be markedly permissive. This is the initial reporting of CCHFV susceptibility and replication patterns in bovine cells and the first report to compare human and animal cell permissiveness in vitro. Further investigations will help to understand the impact of different cell types of various origins on the virus–host interaction.

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