Development of monoclonal antibodies to Rift Valley Fever Virus and their application in antigen detection and indirect immunofluorescence

Mroz, Claudia; Schmidt, Kristina Maria; Reiche, Sven; Groschup, Martin H.; Eiden, Martin

doi:10.1016/j.jim.2018.06.006

Cited by 7 publications

(9 citation statements)

References 30 publications

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“…Vero cells were infected at a multiplicity of infection (MOI) of 1 and, at the time postinfection (pi) indicated, cells were fixed with 4% paraformaldehyde and subjected to indirect immunofluorescence with the anti-NSs monoclonal antibody 5C3A1B12 [20], kindly provided by Dr. Martin Eiden (Friedrich-Loeffler Institute, Riems, Germany) following procedures as described [16]. All the buffers included 0.1% saponin for permeabilization.…”

Section: Immunofluorescencementioning

confidence: 99%

“…P82 is placed within PXXP motif 2 (positions 82 to 85), and changes in two of the four PXXP motifs present in the NSs (motif 1 at positions 29 to 32, and motif 2 at positions 82 to 85) are known to affect the nuclear filamentous arrangement of NSs, with NSs mutants remaining in the cytoplasm [15]. In order to test whether the change carried by our P82L-mutants affected this pattern, infected Vero cells were subjected to indirect immunofluorescence with the anti-NSs monoclonal antibody 5C3AB12 [20]. At 6 hpi, NSs could be detected in the cytoplasm of all infected cells in all cases, but in the nucleus the prototypical fibrillar NSs structures could only be detected after infection with wt rZH548 virus (Figure 3A, left panels).…”

Section: Cellular Localization and Pattern Of Nss In Cells Infected With Rzh548-p82l Mutantsmentioning

confidence: 99%

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The Change P82L in the Rift Valley Fever Virus NSs Protein Confers Attenuation in Mice

Borrego

Moreno

Losa

et al. 2021

Viruses

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Rift Valley fever virus (RVFV) is a mosquito-borne bunyavirus that causes an important disease in ruminants, with great economic losses. The infection can be also transmitted to humans; therefore, it is considered a major threat to both human and animal health. In a previous work, we described a novel RVFV variant selected in cell culture in the presence of the antiviral agent favipiravir that was highly attenuated in vivo. This variant displayed 24 amino acid substitutions in different viral proteins when compared to its parental viral strain, two of them located in the NSs protein that is known to be the major virulence factor of RVFV. By means of a reverse genetics system, in this work we have analyzed the effect that one of these substitutions, P82L, has in viral attenuation in vivo. Rescued viruses carrying this single amino acid change were clearly attenuated in BALB/c mice while their growth in an interferon (IFN)-competent cell line as well as the production of interferon beta (IFN-β) did not seem to be affected. However, the pattern of nuclear NSs accumulation was modified in cells infected with the mutant viruses. These results highlight the key role of the NSs protein in the modulation of viral infectivity.

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Section: Immunofluorescencementioning

confidence: 99%

Section: Cellular Localization and Pattern Of Nss In Cells Infected With Rzh548-p82l Mutantsmentioning

confidence: 99%

The Change P82L in the Rift Valley Fever Virus NSs Protein Confers Attenuation in Mice

Borrego

Moreno

Losa

et al. 2021

Viruses

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“…were fixed with 4% paraformaldehyde. Subsequently, an IIFA to verify RVFV infection of these cells was performed as previously described [ 35 ]. A monoclonal antibody against RVFV NP was used as first antibody.…”

Section: Methodsmentioning

confidence: 99%

Mosquito survey in Mauritania: Detection of Rift Valley fever virus and dengue virus and the determination of feeding patterns

Stoek

Barry²,

Ba³

et al. 2022

PLoS Negl Trop Dis

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In Mauritania, several mosquito-borne viruses have been reported that can cause devastating diseases in animals and humans. However, monitoring data on their occurrence and local distribution are limited. Rift Valley fever virus (RVFV) is an arthropod-borne virus that causes major outbreaks throughout the African continent and the Arabian Peninsula. The first Rift Valley fever (RVF) epidemic in Mauritania occurred in 1987 and since then the country has been affected by recurrent outbreaks of the disease. To gain information on the occurrence of RVFV as well as other mosquito-borne viruses and their vectors in Mauritania, we collected and examined 4,950 mosquitoes, belonging to four genera and 14 species. The mosquitoes were captured during 2018 in the capital Nouakchott and in southern parts of Mauritania. Evidence of RVFV was found in a mosquito pool of female Anopheles pharoensis mosquitoes collected in December on a farm near the Senegal River. At that time, 37.5% of 16 tested Montbéliarde cattle on the farm showed RVFV-specific IgM antibodies. Additionally, we detected IgM antibodies in 10.7% of 28 indigenous cattle that had been sampled on the same farm one month earlier. To obtain information on potential RVFV reservoir hosts, blood meals captured engorged mosquitoes were analyzed. The mosquitoes mainly fed on humans (urban areas) and cattle (rural areas), but also on small ruminants, donkeys, cats, dogs and straw-colored fruit bats. Results of this study demonstrate the circulation of RVFV in Mauritania and thus the need for further research to investigate the distribution of the virus and its vectors. Furthermore, factors that may contribute to its maintenance should be analyzed more closely. In addition, two mosquito pools containing Aedes aegypti and Culex quinquefasciatus mosquitoes showed evidence of dengue virus (DENV) 2 circulation in the city of Rosso. Further studies are therefore needed to also examine DENV circulation in Mauritania.

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“…Vero cells were infected at a MOI of 1 and, at the time post-infection (pi) indicated, cells were fixed with 4% paraformaldehyde and subjected to indirect immunofluorescence with the anti-NSs monoclonal antibody 5C3A1B12 [20] kindly provided by Dr. Martin Eiden (Friedrich-Loeffler Institute. Riems, Germany) following procedures as described [16].…”

Section: Immunofluorescencementioning

confidence: 99%

“…P82 is placed within PXXP motif 2 (positions 82 to 85), and changes in two of the four PXXP motifs present in the NSs (motif 1 at positions 29 to 32, and motif 2 at positions 82 to 85) are known to affect the nuclear filamentous arrangement of NSs, with NSs mutants remaining in the cytoplasm [15]. In order to test whether the change P82L carried by our S279-mutants affected this pattern, infected Vero cells were subjected to indirect immunofluorescence with the anti-NSs monoclonal antibody 5C3AB12 [20]. At 6 h pi, NSs could be detected in the cytoplasm of all infected cells in all cases, but in the nucleus the prototypical fibrillar NSs structures could only be detected after infection with wt rZH548 virus (figure 3A, left panels).…”

Section: Cellular Localization and Pattern Of Nss In Cells Infected With Rzh548-s279-mutantsmentioning

confidence: 99%

The Change P82L in the Rift Valley Fever Virus NSs Protein Confers Attenuation in Mice Not Related with a Type-I IFN Antagonistic Phenotype

Borrego¹,

Moreno²,

N³

et al. 2021

Preprint

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Rift valley fever virus (RVFV) is a mosquito-borne bunyavirus that causes an important disease in ruminants, with great economic losses. The infection can be also transmitted to humans; therefore it is considered a major threat to both human and animal health. In a previous work, we described a novel RVFV variant selected in cell culture in the presence of the antiviral agent favipiravir that was highly attenuated in vivo. This variant displayed 24 amino acid substitutions in different viral proteins when compared to its parental viral strain, two of them located in the NSs protein that is known to be the major virulence factor of RVFV. By means of a reverse genetics system, in this work we have analyzed the effect that one of these substitutions, P82L, has in viral attenuation in vivo. Rescued viruses carrying this single amino acid change were clearly attenuated in BALB/c mice while their growth in an IFN-competent cell line as well as the production of IFN-β did not seem to be affected. However, the pattern of nuclear NSs accumulation was modified in cells infected with the mutant viruses. These results unveil a new RVFV virulence marker highlighting the multiple ways of NSs protein to modulate viral infectivity.

show abstract

Development of monoclonal antibodies to Rift Valley Fever Virus and their application in antigen detection and indirect immunofluorescence

Cited by 7 publications

References 30 publications

The Change P82L in the Rift Valley Fever Virus NSs Protein Confers Attenuation in Mice

The Change P82L in the Rift Valley Fever Virus NSs Protein Confers Attenuation in Mice

Mosquito survey in Mauritania: Detection of Rift Valley fever virus and dengue virus and the determination of feeding patterns

The Change P82L in the Rift Valley Fever Virus NSs Protein Confers Attenuation in Mice Not Related with a Type-I IFN Antagonistic Phenotype

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