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

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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Hepatocyte pathway alterations in response to in vitro Crimean Congo hemorrhagic fever virus infection

Virus Research ◽

10.1016/j.virusres.2013.10.013 ◽

2014 ◽

Vol 179 ◽

pp. 187-203 ◽

Cited By ~ 15

Author(s):

Christophe Fraisier ◽

Raquel Rodrigues ◽

Vinh Vu Hai ◽

Maya Belghazi ◽

Stéphanie Bourdon ◽

...

Keyword(s):

Virus Infection ◽

Hemorrhagic Fever ◽

Fever Virus ◽

Crimean Congo Hemorrhagic Fever ◽

Hemorrhagic Fever Virus

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Small interfering RNAs are highly effective inhibitors regarding Crimean-Congo hemorrhagic fever virus replication in vitro

10.1101/2020.05.13.093047 ◽

2020 ◽

Author(s):

Fanni Földes ◽

Mónika Madai ◽

Henrietta Papp ◽

Gábor Kemenesi ◽

Brigitta Zana ◽

...

Keyword(s):

Rna Interference ◽

Hemorrhagic Fever ◽

Fever Virus ◽

World Health ◽

Dependent Manner ◽

Small Interfering Rnas ◽

Crimean Congo Hemorrhagic Fever ◽

Hemorrhagic Fever Virus ◽

Antiviral Therapies

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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The impact of Crimean-Congo hemorrhagic fever virus on public health

Antiviral Research ◽

10.1016/j.antiviral.2013.02.007 ◽

2013 ◽

Vol 98 (2) ◽

pp. 248-260 ◽

Cited By ~ 71

Author(s):

Marc Mertens ◽

Katja Schmidt ◽

Aykut Ozkul ◽

Martin H. Groschup

Keyword(s):

Public Health ◽

Hemorrhagic Fever ◽

Fever Virus ◽

Crimean Congo Hemorrhagic Fever ◽

Hemorrhagic Fever Virus ◽

The Impact

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

PLoS ONE ◽

10.1371/journal.pone.0116816 ◽

2015 ◽

Vol 10 (2) ◽

pp. e0116816 ◽

Cited By ~ 8

Author(s):

Licia Bordi ◽

Eleonora Lalle ◽

Claudia Caglioti ◽

Damiano Travaglini ◽

Daniele Lapa ◽

...

Keyword(s):

Antiviral Activity ◽

Hemorrhagic Fever ◽

Fever Virus ◽

Crimean Congo Hemorrhagic Fever ◽

Hemorrhagic Fever Virus ◽

Ifn Alpha ◽

Ifn Lambda

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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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Crimean-Congo hemorrhagic fever virus antibody prevalence in Mauritanian livestock (cattle, goats, sheep and camels) is stratified by the animal’s age

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0009228 ◽

2021 ◽

Vol 15 (4) ◽

pp. e0009228

Author(s):

Ansgar Schulz ◽

Yahya Barry ◽

Franziska Stoek ◽

Aliou Ba ◽

Jana Schulz ◽

...

Keyword(s):

Clinical Symptoms ◽

Small Ruminants ◽

Hemorrhagic Fever ◽

Immunofluorescence Assay ◽

Fever Virus ◽

Crimean Congo Hemorrhagic Fever ◽

Prevalence Studies ◽

Hemorrhagic Fever Virus ◽

Generalized Linear Mixed Effects ◽

The Impact

Crimean-Congo hemorrhagic fever virus (CCHFV) is one of the most widespread zoonotic arthropod-borne viruses in many parts of Africa, Europe and Asia. It belongs to the family of Nairoviridae in the genus of Orthonairovirus. The main reservoir and vector are ticks of the genus Hyalomma. Livestock animals (such as cattle, small ruminants and camels) develop a viremias lasting up to two weeks with absence of clinical symptoms, followed by seroconversion. This study was carried out to assess risk factors that affect seroprevalence rates in different species. In total, 928 livestock animal samples (cattle = 201; sheep = 247; goats = 233; camels = 247) from 11 out of 13 regions in Mauritania were assayed for CCHFV-specific immunoglobulin G (IgG) antibodies using enzyme-linked immunosorbent assays (ELISA) (including a novel indirect camel-IgG-specific CCHFV ELISA). Inconclusive results were resolved by an immunofluorescence assay (IFA). A generalized linear mixed-effects model (GLMM) was used to draw conclusions about the impact of certain factors (age, species, sex and region) which might have influenced the CCHFV antibody status of surveyed animals. In goats and sheep, about 15% of the animals were seropositive, whereas in cattle (69%) and camels (81%), the prevalence rate was significantly higher. On average, cattle and camels were up to twice to four times older than small ruminants. Interestingly, the seroprevalence in all species was directly linked to the age of the animals, i.e. older animals had significantly higher seroprevalence rates than younger animals. The highest CCHFV seroprevalence in Mauritania was found in camels and cattle, followed by small ruminants. The large proportion of positive animals in cattle and camels might be explained by the high ages of the animals. Future CCHFV prevalence studies should at least consider the age of surveyed animals in order to avoid misinterpretations.

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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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