scholarly journals Identification of a Common Conformational Epitope on the Glycoprotein E2 of Classical Swine Fever Virus and Border Disease Virus

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1655
Author(s):  
Yu-Liang Huang ◽  
Denise Meyer ◽  
Alexander Postel ◽  
Kuo-Jung Tsai ◽  
Hsin-Meng Liu ◽  
...  
Keyword(s):  
Disease Virus ◽  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Conformational Epitope ◽  
Cross Reactivity ◽  
Fever Virus ◽  
Site Directed Mutagenesis ◽  
Border Disease Virus ◽  
Diarrhea Virus ◽  
Border Disease

Classical swine fever virus (CSFV) shares high structural and antigenic homology with bovine viral diarrhea virus (BVDV) and border disease virus (BDV). Because all three viruses can infect swine and elicit cross-reactive antibodies, it is necessary to differentiate among them with regard to serological diagnosis of classical swine fever. To understand the mechanism of cross-reactivity, it is important to define common or specific epitopes of these viruses. For this purpose, epitope mapping of six monoclonal antibodies (mAbs) was performed using recombinant expressed antigenic domains of CSFV and BDV E2 proteins. One CSFV-specific conformational epitope and one CSFV and BDV common epitope within domain B/C of E2 were identified. Site-directed mutagenesis confirmed that residues G725 and V738/I738 of the CSFV-specific epitope and P709/L709 and E713 of the second epitope are important for mAbs binding. Infection of CSFV in porcine cells was significantly reduced after pre-incubation of the cells with the domain B/C of E2 or after pre-incubation of CSFV with the mAbs detecting domain B/C. 3D structural modeling suggested that both epitopes are exposed on the surface of E2. Based on this, the identified epitopes represent a potential target for virus neutralization and might be involved in the early steps of CSFV infection.

scholarly journals Influence of NS5A protein of classical swine fever virus (CSFV) on CSFV internal ribosome entry site-dependent translation

2009 ◽  
Vol 90 (12) ◽  
pp. 2923-2928 ◽  
Author(s):  
Ming Xiao ◽  
Yujing Wang ◽  
Zailing Zhu ◽  
Jialin Yu ◽  
Lingzhu Wan ◽  
...  
Keyword(s):  
Classical Swine Fever Virus ◽  
Internal Ribosome Entry Site ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Border Disease Virus ◽  
Dependent Manner ◽  
Entry Site ◽  
Internal Ribosome Entry ◽  
Diarrhea Virus ◽  
Border Disease

An internal ribosome entry site (IRES) present in the 5′ untranslated region (UTR) promotes translation of classical swine fever virus (CSFV) genomes. Using an in vitro system with monocistronic reporter RNA containing the CSFV 5′UTR, this study found that CSFV NS5A decreased CSFV IRES-mediated translation in a dose-dependent manner. Deletion analysis showed that the region responsible for repressing CSFV IRES activity might cover aa  390–414, located in the C-terminal half of CSFV NS5A. Triple and single alanine-scanning mutagenesis revealed that the inhibitory effect on CSFV IRES-directed translation mapped to the K399, T401, E406 and L413 residues of NS5A. These important amino acids were also found to be present in the NS5A proteins of bovine viral diarrhea virus (BVDV)-1, BVDV-2, border disease virus and hepatitis C virus, indicating that NS5A may play an important role in the switch from translation to replication in these viruses.

scholarly journals Serum neutralization as a differential serological test for classical swine fever virus and other pestivirus infections

1999 ◽  
Vol 51 (5) ◽  
pp. 403-408 ◽  
Author(s):  
J.C.M. Paredes ◽  
E.A.S. Oliveira ◽  
L.G. Oliveira ◽  
J.C.A. Rosa ◽  
P.M. Roehe
Keyword(s):  
Classical Swine Fever Virus ◽  
Serological Test ◽  
Sampling Strategy ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Border Disease Virus ◽  
Serum Samples ◽  
Diarrhea Virus ◽  
Serum Neutralization ◽  
Border Disease

Serum neutralization tests (SN) were performed against classical swine fever virus (CSFV), bovine viral diarrhea virus (BVDV) and border disease virus (BDV) on samples of swine serum collected for screening of antibodies to CSFV, in order to determine the SN value as a differential serological test. Ninety-nine sera out of a sample of 16,664 were positive for antibodies to pestiviruses in an ELISA test which did not distinguish antibodies to different pestiviruses. When submitted to SN, 81 sera were positive for CSFV antibodies only. In 17 sera, crossreactive antibodies to either CSFV, BVDV or BDV were detected. In most of these sera (13 out of 17) the differences between SN titres against the three viruses were not sufficient to estimate which was the most likely antibody-inducing virus. It was concluded that, for the SN to be useful in such differentiation, it is essential to examine a sample which must include a representative number of sera from the same farm where suspect animals were detected. When isolated serum samples are examined, such as those obtained with the sampling strategy adopted here, the SN may give rise to inconclusive results.

scholarly journals Comparative Analysis of Tunisian Sheep-Like Virus, Bungowannah Virus and Border Disease Virus Infection in the Porcine Host

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1539
Author(s):  
Denise Meyer ◽  
Alexander Postel ◽  
Anastasia Wiedemann ◽  
Gökce Nur Cagatay ◽  
Sara Ciulli ◽  
...  
Keyword(s):  
Disease Virus ◽  
Classical Swine Fever ◽  
Cross Reactivity ◽  
Border Disease Virus ◽  
Domestic Pigs ◽  
Diarrhea Virus ◽  
Antigenic Relatedness ◽  
Border Disease ◽  
Antibody Assays ◽  
Viral Diarrhea Virus

Apart from the established pestivirus species Pestivirus A to Pestivirus K novel species emerged. Pigs represent not only hosts for porcine pestiviruses, but are also susceptible to bovine viral diarrhea virus, border disease virus (BDV) and other ruminant pestiviruses. The present study focused on the characterization of the ovine Tunisian sheep-like virus (TSV) as well as Bungowannah virus (BuPV) and BDV strain Frijters, which were isolated from pigs. For this purpose, we performed genetic characterization based on complete coding sequences, studies on virus replication in cell culture and in domestic pigs, and cross-neutralization assays using experimentally derived sera. TSV forms a distinct phylogenetic group more closely related to Pestivirus C (classical swine fever virus, CSFV) than to Pestivirus D (BDV). In contrast to BDV and BuPV, TSV replicates by far more efficiently on ovine than on porcine cells. Nevertheless, pigs were susceptible to TSV. As a consequence of close antigenic relatedness of TSV to CSFV, cross-reactivity was detected in CSFV-specific antibody assays. In conclusion, TSV is genetically closely related to CSFV and can replicate in domestic pigs. Due to close antigenic relatedness, field infections of pigs with TSV and other ruminant pestiviruses can interfere with serological diagnosis of classical swine fever.

scholarly journals Development of a High-Throughput Serum Neutralization Test Using Recombinant Pestiviruses Possessing a Small Reporter Tag

Pathogens ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 188 ◽  
Author(s):  
Madoka Tetsuo ◽  
Keita Matsuno ◽  
Tomokazu Tamura ◽  
Takasuke Fukuhara ◽  
Taksoo Kim ◽  
...  
Keyword(s):  
High Throughput ◽  
Neutralization Test ◽  
Classical Swine Fever ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Border Disease Virus ◽  
Diarrhea Virus ◽  
Serum Neutralization ◽  
Border Disease ◽  
Serum Neutralization Test

A serum neutralization test (SNT) is an essential method for the serological diagnosis of pestivirus infections, including classical swine fever, because of the cross reactivity of antibodies against pestiviruses and the non-quantitative properties of antibodies in an enzyme-linked immunosorbent assay. In conventional SNTs, an immunoperoxidase assay or observation of cytopathic effect after incubation for 3 to 7 days is needed to determine the SNT titer, which requires labor-intensive or time-consuming procedures. Therefore, a new SNT, based on the luciferase system and using classical swine fever virus, bovine viral diarrhea virus, and border disease virus possessing the 11-amino-acid subunit derived from NanoLuc luciferase was developed and evaluated; this approach enabled the rapid and easy determination of the SNT titer using a luminometer. In the new method, SNT titers can be determined tentatively at 2 days post-infection (dpi) and are comparable to those obtained by conventional SNTs at 3 or 4 dpi. In conclusion, the luciferase-based SNT can replace conventional SNTs as a high-throughput antibody test for pestivirus infections.

scholarly journals Complete Genomic Sequence of Border Disease Virus, a Pestivirus from Sheep

1998 ◽  
Vol 72 (6) ◽  
pp. 5165-5173 ◽  
Author(s):  
Paul Becher ◽  
Michaela Orlich ◽  
Heinz-Jürgen Thiel
Keyword(s):  
Serine Protease ◽  
Disease Virus ◽  
Genomic Sequence ◽  
Classical Swine Fever ◽  
Border Disease Virus ◽  
Sequence Motif ◽  
Reading Frame ◽  
Diarrhea Virus ◽  
Expression Studies ◽  
Border Disease

ABSTRACT The genus Pestivirus of the familyFlaviviridae comprises three established species, namely, bovine viral diarrhea virus (BVDV), classical swine fever virus (CSFV), and border disease virus from sheep (BDV). In this study, we report the first complete nucleotide sequence of BDV, that of strain X818. The genome is 12,333 nucleotides long and contains one long open reading frame encoding 3,895 amino acids. The 5′ noncoding region (NCR) of BDV X818 consists of 372 nucleotides and is thus similar in length to the 5′ NCR reported for other pestiviruses. The 3′ NCR of X818 is 273 nucleotides long and thereby at least 32 nucleotides longer than the 3′ NCR of pestiviruses analyzed thus far. Within the 3′ NCR of BDV X818, the sequence motif TATTTATTTA was identified at four locations. The same repeat was found at two or three locations within the 3′ NCR of different CSFV isolates but was absent in the 3′ NCR of BVDV. Analysis of five additional BDV strains showed that the 3′ NCR sequences are highly conserved within this species. Comparison of the deduced amino acid sequence of X818 with the ones of other pestiviruses allowed the prediction of polyprotein cleavage sites which were conserved with regard to the structural proteins. It has been reported for two BVDV strains that cleavage at the nonstructural (NS) protein sites 3/4A, 4A/4B, 4B/5A, and 5A/5B is mediated by the NS3 serine protease and for each site a conserved leucine was found at the P1 position followed by either serine or alanine at P1′ (N. Tautz, K. Elbers, D. Stoll, G. Meyers, and H.-J. Thiel, J. Virol. 71:5415–5422, 1997; J. Xu, E. Mendez, P. R. Caron, C. Lin, M. A. Murcko, M. S. Collett, and C. M. Rice, J. Virol. 71:5312–5322). Interestingly, P1′ of the predicted NS5A/5B cleavage site of BDV is represented by an asparagine residue. Transient expression studies demonstrated that this unusual NS5A/5B processing site is efficiently cleaved by the NS3 serine protease of BDV.

scholarly journals Global Distribution and Genetic Heterogeneity of Border Disease Virus

Viruses ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 950
Author(s):  
Cecilia Righi ◽  
Stefano Petrini ◽  
Ilaria Pierini ◽  
Monica Giammarioli ◽  
Gian Mario De Mia
Keyword(s):  
Disease Virus ◽  
Significant Risk ◽  
Small Ruminants ◽  
Control Measures ◽  
Interspecies Transmission ◽  
Economic Losses ◽  
Border Disease Virus ◽  
Diarrhea Virus ◽  
Geographical Regions ◽  
Border Disease

Border disease virus (BDV) belongs to the genus Pestivirus of the family Flaviviridae. Interspecies transmission of BDV between sheep, cattle, and pigs occurs regularly, sometimes making diagnosis a challenge. BDV can yield substantial economic losses, including prenatal and postnatal infections in lambs, which are the primary source of infection and maintenance of the virus in the population. Since BDV is antigenically and genetically related to bovine viral diarrhea virus (BVDV), it might pose a significant risk to cattle, influencing BVDV eradication campaigns. Similarly, the presence of BDV in swine herds due to pestivirus spillover between small ruminants and pigs might cause uncertainty in classical swine fever virus (CSFV) diagnostics. Therefore, knowledge of BDV epidemiology in different geographical regions will help prevent its spread and optimize control measures. Previous epidemiological studies have shown that various BDV genotypes are predominant in different countries. This review provides an overview of the spread of BDV world-wide in different host species.

scholarly journals Porcine complement regulatory protein CD46 is a major receptor for atypical porcine pestivirus but not for classical swine fever virus

2021 ◽  
Author(s):  
Gökce Nur Cagatay ◽  
Aleksandra Antos ◽  
Oliver Suckstorff ◽  
Olaf Isken ◽  
Norbert Tautz ◽  
...  
Keyword(s):  
Classical Swine Fever Virus ◽  
Regulatory Protein ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Bovine Viral Diarrhea ◽  
Complement Regulatory Protein ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Viral Diarrhea Virus ◽  
Entry Mechanism

Pestiviruses such as bovine viral diarrhea virus (BVDV) and classical swine fever virus (CSFV) belong to the family Flaviviridae and represent pathogens of outstanding veterinary relevance. Pestiviruses enter cells via receptor-mediated endocytosis. For entry in bovine cells, complement regulatory protein CD46bov serves as cellular receptor for BVDV. In this study, the role of porcine CD46pig in cellular entry was investigated for the recently discovered atypical porcine pestivirus (APPV), CSFV, and Bungowannah virus (BuPV) in order to elucidate the observed differences in host cell tropism. A cell culture adapted APPV variant, which shows enhanced viral replication in vitro, was generated and demonstrated a strict tropism of APPV for porcine cells. One of the porcine cell lines displayed areas of CD46pig expressing and areas of non-expressing cells and one single cell line revealed not to express any CD46pig, respectively. The CD46pig deficient porcine lymphoma cells, known to facilitate CSFV replication, was the only porcine cell line non-permissive to APPV, indicating a significant difference in the entry mechanism of APPV and CSFV. Infection experiments with a set of genetically engineered CD46pig knockout cells confirmed that CD46pig is a major receptor of APPV as CD46bov is for BVDV. In contrast, it is apparently not an essential determinant in host cell entry of other porcine pestiviruses such as CSFV and BuPV. Existence of a CD46pig independent entry mechanism illustrates that the pestiviral entry process is more diverse than previously recognized. IMPORTANCE Pestiviruses comprise animal pathogens such as classical swine fever virus (CSFV) and bovine viral diarrhea virus (BVDV) that cause notifiable diseases with great economic impact. Several additional pestivirus species affecting animal health were recently identified, including atypical porcine pestivirus (APPV). APPV is associated with health problems in piglets and highly abundant in pig populations worldwide. Complement control protein CD46 serves as a receptor for diverse bacterial and viral pathogens, including particular adenoviruses, herpesviruses, measles virus (MeV), and BVDV. Porcine CD46 (CD46pig) was suggested to be a major receptor for CSFV. Here, we identified remarkable differences in relevance of CD46pig during entry of porcine pestiviruses. Resembling BVDV, efficient APPV infection in cell culture depends on CD46pig, while other porcine pestiviruses can efficiently enter and infect cells in absence of CD46pig. Thus, the study provides insights into the entry process of these pathogens and may help to understand differences in their biology.

scholarly journals Determination of Critical Requirements for Classical Swine Fever Virus NS2-3-Independent Virion Formation

2019 ◽  
Vol 93 (18) ◽  
Author(s):  
D. Dubrau ◽  
S. Schwindt ◽  
O. Klemens ◽  
H. Bischoff ◽  
N. Tautz
Keyword(s):  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Rna Replication ◽  
Fever Virus ◽  
Genome Packaging ◽  
Gain Of Function ◽  
Diarrhea Virus ◽  
Virion Production ◽  
Virion Morphogenesis ◽  
Virion Formation

ABSTRACTFor members of theFlaviviridae, it is known that, besides the structural proteins, nonstructural (NS) proteins also play a critical role in virion formation. Pestiviruses, such as bovine viral diarrhea virus (BVDV), rely on uncleaved NS2-3 for virion formation, while its cleavage product, NS3, is selectively active in RNA replication. This dogma was recently challenged by the selection of gain-of-function mutations in NS2 and NS3 which allowed virion formation in the absence of uncleaved NS2-3 in BVDV type 1 (BVDV-1) variants encoding either a ubiquitin (Ubi) (NS2-Ubi-NS3) or an internal ribosome entry site (IRES) (NS2-IRES-NS3) between NS2 and NS3. To determine whether the ability to adapt to NS2-3-independent virion morphogenesis is conserved among pestiviruses, we studied the corresponding NS2 and NS3 mutations (2/T444-V and 3/M132-A) in classical swine fever virus (CSFV). We observed that these mutations were capable of restoring low-level NS2-3-independent virion formation only for CSFV NS2-Ubi-NS3. Interestingly, a second NS2 mutation (V439-D), identified by selection, was essential for high-titer virion production. Similar to previous findings for BVDV-1, these mutations in NS2 and NS3 allowed for low-titer virion production only in CSFV NS2-IRES-NS3. For efficient virion morphogenesis, additional exchanges in NS4A (A48-T) and NS5B (D280-G) were required, indicating that these proteins cooperate in NS2-3-independent virion formation. Interestingly, both NS5B mutations, selected independently for NS2-IRES-NS3 variants of BVDV-1 and CSFV, are located in the fingertip region of the viral RNA-dependent RNA polymerase, classifying this structural element as a novel determinant for pestiviral NS2-3-independent virion formation. Together, these findings will stimulate further mechanistic studies on the genome packaging of pestiviruses.IMPORTANCEForFlaviviridaemembers, the nonstructural proteins are essential for virion formation and thus exert a dual role in RNA replication and virion morphogenesis. However, it remains unclear how these proteins are functionalized for either process. In wild-type pestiviruses, the NS3/4A complex is selectively active in RNA replication, while NS2-3/4A is essential for virion formation. Mutations recently identified in BVDV-1 rendered NS3/4A capable of supporting NS2-3-independent virion morphogenesis. A comparison of NS3/4A complexes incapable/capable of supporting virion morphogenesis revealed that changes in NS3/NS4A surface interactions are decisive for the gain of function. However, so far, the role of the NS2 mutations as well as the accessory mutations additionally required in the NS2-IRES-NS3 virus variant has not been clarified. To unravel the course of genome packaging, the additional sets of mutations obtained for a second pestivirus species (CSFV) are of significant importance to develop mechanistic models for this complex process.

scholarly journals Characterization of Helper Virus-Independent Cytopathogenic Classical Swine Fever Virus Generated by an In Vivo RNA Recombination System

2005 ◽  
Vol 79 (4) ◽  
pp. 2440-2448 ◽  
Author(s):  
Andreas Gallei ◽  
Till Rümenapf ◽  
Heinz-Jürgen Thiel ◽  
Paul Becher
Keyword(s):  
Classical Swine Fever Virus ◽  
Nonstructural Protein ◽  
Classical Swine Fever ◽  
Helper Virus ◽  
Fever Virus ◽  
Tissue Culture Cell ◽  
Rna Recombination ◽  
Diarrhea Virus ◽  
Recombination System

ABSTRACT Molecular analyses revealed that most cytopathogenic (cp) pestivirus strains evolve from noncytopathogenic (noncp) viruses by nonhomologous RNA recombination. In contrast to bovine viral diarrhea virus (BVDV), cp classical swine fever virus (CSFV) field isolates were rarely detected and always represented helper virus-dependent subgenomes. To investigate RNA recombination in more detail, we recently established an in vivo system allowing the efficient generation of recombinant cp BVDV strains in cell culture after transfecting a synthetic subgenomic and nonreplicatable transcript into cells being infected with noncp BVDV (A. Gallei, A. Pankraz, H.-J. Thiel, and P. Becher, J. Virol. 78:6271-6281, 2004). Using an analogous approach, the first helper virus-independent cp CSFV strain (CP G1) has now been generated by RNA recombination. Accordingly, this study demonstrates the applicability of RNA recombination for designing new viral RNA genomes. The genomic RNA of CP G1 has a calculated size of 18.139 kb, almost 6 kb larger than all previously described CSFV genomes. It contains cellular sequences encoding a polyubiquitin fragment directly upstream of the nonstructural protein NS3 coding gene together with a duplication of viral sequences. CP G1 induces a cytopathic effect on different tissue culture cell lines from pigs and cattle. Subsequent analyses addressed growth kinetics, expression of NS3, and genetic stability of CP G1.

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