The Experts below are selected from a list of 4989 Experts worldwide ranked by ideXlab platform
Huaji Qiu - One of the best experts on this subject based on the ideXlab platform.
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isolation and characterization of a moderately virulent Classical Swine Fever virus emerging in china
Transboundary and Emerging Diseases, 2017Co-Authors: Yuzi Luo, Yuan Sun, Yonggang Liu, Jianlin Lei, Shuili Xia, Yimin Wang, Lina Shao, Xingyu Meng, Mo Zhou, Huaji QiuAbstract:Classical Swine Fever (CSF) is a devastating infectious disease of pigs caused by Classical Swine Fever virus (CSFV). In China, CSF has been under control owing to extensive vaccination with the lapinized attenuated vaccine (C-strain) since 1950s, despite sporadic or endemic in many regions. However, recently, CSF outbreaks occurred in a large number of Swine herds in China. Here, we isolated 15 CSFV strains from diverse C-strain-vaccinated pig farms in China and characterized the genetic variations and antigenicity of the new isolates. The new strains showed unique variations in the E2 protein and were clustered to the subgenotype 2.1d of CSFV recently emerging in China in the phylogenetic tree. Cross-neutralization test showed that the neutralizing titres of porcine anti-C-strain sera against the new isolates were substantially lower than those against both the highly virulent Shimen strain and the subgenotype 2.1b strains that were isolated in China in 2006 and 2009, respectively. In addition, experimental animal infection showed that the HLJZZ2014 strain-infected pigs displayed lower mortality and less severe clinical signs and pathological changes compared with the Shimen strain-infected pigs. The HLJZZ2014 strain was defined to be moderately virulent based on a previously established assessment system for CSFV virulence evaluation, and the virus shedding and the viral load in various tissues of the CSFV HLJZZ2014 strain-infected pigs were significantly lower than those of the Shimen strain-infected pigs. Taken together, the subgenotype 2.1d isolate of CSFV is a moderately virulent strain with molecular variations and antigenic alterations.
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interferon inducible oligoadenylate synthetase like protein acts as an antiviral effector against Classical Swine Fever virus via the mda5 mediated type i interferon signaling pathway
Journal of Virology, 2017Co-Authors: Yuexiu Zhang, Yuzi Luo, Yuan Sun, Jinghan Wang, Lingkai Zhang, Qian Yang, Huaji QiuAbstract:Classical Swine Fever virus (CSFV) is the causative agent of Classical Swine Fever (CSF), which poses a serious threat to the global pig industry. Interferons (IFNs) and IFN-stimulated genes (ISGs) play a key role in host antiviral defense. We have previously screened the porcine 2'-5'-oligoadenylate synthetase-like protein (pOASL) as a potential anti-CSFV ISG using a reporter CSFV. This study aimed to clarify the underlying antiviral mechanism of pOASL against CSFV. We confirmed that CSFV replication was significantly suppressed in lentivirus-delivered, pOASL-overexpressing PK-15 cells, whereas silencing the expression of endogenous pOASL by small interfering RNAs markedly enhanced CSFV growth. In addition, the transcriptional level of pOASL was upregulated both in vitro and in vivo upon CSFV infection. Interestingly, the anti-CSFV effects of pOASL are independent of the canonical RNase L pathway but depend on the activation of the type I IFN response. Glutathione S-transferase pulldown and coimmunoprecipitation assays revealed that pOASL interacts with MDA5, a double-stranded RNA sensor, and further enhances MDA5-mediated type I IFN signaling. Moreover, we showed that pOASL exerts anti-CSFV effects in an MDA5-dependent manner. In conclusion, pOASL suppresses CSFV replication via the MDA5-mediated type I IFN-signaling pathway.IMPORTANCE The host innate immune response plays an important role in mounting the initial resistance to viral infection. Here, we identify the porcine 2'-5'-oligoadenylate synthetase-like protein (pOASL) as an interferon (IFN)-stimulated gene (ISG) against Classical Swine Fever virus (CSFV). We demonstrate that the anti-CSFV effects of pOASL depend on the activation of type I IFN response. In addition, we show that pOASL, as an MDA5-interacting protein, is a coactivator of MDA5-mediated IFN induction to exert anti-CSFV actions. This work will be beneficial to the development of novel anti-CSFV strategies by targeting pOASL.
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guanylate binding protein 1 an interferon induced gtpase exerts an antiviral activity against Classical Swine Fever virus depending on its gtpase activity
Journal of Virology, 2016Co-Authors: Jinghan Wang, Xiao Wang, Yuzi Luo, Yuan Sun, Shuili Xia, Lingkai Zhang, Qian Yang, Yan Zhu, Muhammad Munir, Huaji QiuAbstract:UNLABELLED: Many viruses trigger the type I interferon (IFN) pathway upon infection, resulting in the transcription of hundreds of interferon-stimulated genes (ISGs), which define the antiviral state of the host. Classical Swine Fever virus (CSFV) is the causative agent of Classical Swine Fever (CSF), a highly contagious viral disease endangering the pig industry in many countries. However, anti-CSFV ISGs are poorly documented. Here we screened 20 ISGs that are commonly induced by type I IFNs against CSFV in lentivirus-delivered cell lines, resulting in the identification of guanylate-binding protein 1 (GBP1) as a potent anti-CSFV ISG. We observed that overexpression of GBP1, an IFN-induced GTPase, remarkably suppressed CSFV replication, whereas knockdown of endogenous GBP1 expression by small interfering RNAs significantly promoted CSFV growth. Furthermore, we demonstrated that GBP1 acted mainly on the early phase of CSFV replication and inhibited the translation efficiency of the internal ribosome entry site of CSFV. In addition, we found that GBP1 was upregulated at the transcriptional level in CSFV-infected PK-15 cells and in various organs of CSFV-infected pigs. Coimmunoprecipitation and glutathione S-transferase (GST) pulldown assays revealed that GBP1 interacted with the NS5A protein of CSFV, and this interaction was mapped in the N-terminal globular GTPase domain of GBP1. Interestingly, the K51 of GBP1, which is crucial for its GTPase activity, was essential for the inhibition of CSFV replication. We showed further that the NS5A-GBP1 interaction inhibited GTPase activity, which was critical for its antiviral effect. Taking our findings together, GBP1 is an anti-CSFV ISG whose action depends on its GTPase activity. IMPORTANCE: Classical Swine Fever virus (CSFV) is the causative agent of Classical Swine Fever (CSF), an economically important viral disease affecting the pig industry in many countries. To date, only a few host restriction factors against CSFV, including interferon-stimulated genes (ISGs), have been characterized. Using a minilibrary of porcine ISGs, we identify porcine guanylate-binding protein 1 (GBP1) as a potent antiviral ISG against CSFV. We further show that the anti-CSFV action of GBP1 depends on its GTPase activity. The K51 of GBP1, critical for its GTPase activity, is essential for the antiviral action of GBP1 against CSFV replication, and the binding of the NS5A protein to GBP1 antagonizes the GTPase activity and thus the antiviral effect. This study will facilitate the development of anti-CSFV therapeutic agents by targeting host factors and may provide a new strategy for the control of CSF.
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the laminin receptor is a cellular attachment receptor for Classical Swine Fever virus
Journal of Virology, 2015Co-Authors: Jianing Chen, Liang Shen, Hong Dong, Xiao Wang, Yuzi Luo, Yuan Sun, Huaji QiuAbstract:UNLABELLED Classical Swine Fever virus (CSFV) is the causative agent of Classical Swine Fever (CSF), a highly contagious, economically important viral disease in many countries. The E(rns) and E2 envelope glycoproteins are responsible for the binding to and entry into the host cell by CSFV. To date, only one cellular receptor, heparan sulfate (HS), has been identified as being involved in CSFV attachment. HS is also present on the surface of various cells that are nonpermissive to CSFV. Hence, there must be another receptor(s) that has been unidentified to date. In this study, we used a set of small interfering RNAs (siRNAs) against a number of porcine cell membrane protein genes to screen cellular proteins involved in CSFV infection. This approach resulted in the identification of several proteins, and of these, the laminin receptor (LamR) has been demonstrated to be a cellular receptor for several viruses. Confocal analysis showed that LamR is colocalized with CSFV virions on the membrane, and a coimmunoprecipitation assay indicated that LamR interacts with the CSFV E(rns) protein. In inhibition assays, anti-LamR antibodies, soluble laminin, or LamR protein significantly inhibited CSFV infection in a dose-dependent manner. Transduction of PK-15 cells with a recombinant lentivirus expressing LamR yielded higher viral titers. Moreover, an attachment assay demonstrated that LamR functions during virus attachment. We also demonstrate that LamR acts as an alternative attachment receptor, especially in SK6 cells. These results indicate that LamR is a cellular attachment receptor for CSFV. IMPORTANCE Classical Swine Fever virus (CSFV) is the causative agent of Classical Swine Fever (CSF), an economically important viral disease affecting the pig industry in many countries. To date, only heparan sulfate (HS) has been identified to be an attachment receptor for CSFV. Here, using RNA interference screening with small interfering RNAs (siRNAs) against a number of porcine membrane protein genes, we identified the laminin receptor (LamR) to be another attachment receptor. We demonstrate the involvement of LamR together with HS in virus attachment, and we elucidate the relationship between LamR and HS. LamR also serves as an attachment receptor for many viral pathogens, including dengue virus, a fatal human flavivirus. The study will help to enhance our understanding of the life cycle of flaviviruses and the development of antiviral strategies for flaviviruses.
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specific ligands for Classical Swine Fever virus screened from landscape phage display library
Antiviral Research, 2014Co-Authors: Yuzi Luo, Huaji Qiu, Long Yin, Bo Liang, Fei Wang, Valery A Petrenko, Aihua LiuAbstract:Classical Swine Fever (CSF) is a devastating infectious disease caused by Classical Swine Fever virus (CSFV). The screening of CSFV-specific ligands is of great significance for diagnosis and treatment of CSF. Affinity selection from random peptide libraries is an efficient approach to discover ligands with high stability and specificity. Here, we screened phage ligands for the CSFV E2 protein from f8/8 landscape phage display library by biopanning and obtained four phage clones specific for the E2 protein of CSFV. Viral blocking assays indicated that the phage clone displaying the octapeptide sequence DRATSSNA remarkably inhibited the CSFV replication in PK-15 cells at a titer of 10(10) transduction units, as evidenced by significantly decreased viral RNA copies and viral titers. The phage-displayed E2-binding peptides have the potential to be developed as antivirals for CSF. (C) 2014 Elsevier B.V. All rights reserved.
H.j. Thiel - One of the best experts on this subject based on the ideXlab platform.
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Classical Swine Fever virus independent induction of protective immunity by two structural glycoproteins
Journal of Virology, 1995Co-Authors: Matthias Konig, T Lengsfeld, T Pauly, R Stark, H.j. ThielAbstract:To study which proteins of Classical Swine Fever virus (CSFV) are able to confer protective immunity in Swine, N-terminal autoprotease, viral core protein, and the three structural glycoproteins were expressed via vaccinia virus recombinants (VVR). CSFV proteins synthesized in cells infected with VVR showed migration characteristics on sodium dodecyl sulfate gels identical to those of their respective CSFV counterparts. Apparently authentic dimerization of the recombinant glycoproteins was observed. The glycoproteins E0 and E2 were detected on the surfaces of VVR-infected cells. In protection experiments, Swine were immunized with the different VVR, and the generation of humoral immune response was monitored. Only animals vaccinated with VVR expressing E0 and/or E2 resisted a lethal challenge infection with CSFV. Glycoprotein E0 represents a second determinant for the induction of protective immunity against Classical Swine Fever.
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Classical Swine Fever virus independent induction of protective immunity by two structural glycoproteins
Journal of Virology, 1995Co-Authors: Matthias Konig, T Lengsfeld, T Pauly, R Stark, H.j. ThielAbstract:Classical Swine Fever (CSF), a highly contagious infection of Swine, is caused by a small enveloped RNA virus named Classical Swine Fever virus (CSFV) that is classified as belonging to the genus Pestivirus of the family Flaviviridae (36). The vast economic importance of CSF initiated various efforts to extinguish the disease. Vaccination with attenuated live virus is safe and effective but interferes with serodiagnosis and the maintenance of a virus-free status. In order to avoid trade restrictions, eradication programs are mostly based on the destruction of infected and serologically positive animals. In spite of continued attempts to control CSF, outbreaks have occurred intermittently in several European countries. In some areas, eradication is complicated by the prevalence of CSFV in the
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cytopathogenicity of Classical Swine Fever virus caused by defective interfering particles
Journal of Virology, 1995Co-Authors: G Meyers, H.j. ThielAbstract:For three independent cytopathogenic isolates of Classical Swine Fever virus, defective RNAs were found in infected cells in addition to full-length viral genomes. These RNAs represent the genomes of typical defective interfering (DI) particles because of strict dependence on a complementing helper virus and interference with the replication of the helper virus. Analysis of the DI genomes revealed internal deletions of 4,764 nucleotides encompassing the complete structural protein-coding region of the virus and two flanking nonstructural genes. Plaque isolation and RNA transfection experiments showed that the DI particles are responsible for the cytopathic effect caused by these Classical Swine Fever virus isolates.
Long Yin - One of the best experts on this subject based on the ideXlab platform.
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specific ligands for Classical Swine Fever virus screened from landscape phage display library
Antiviral Research, 2014Co-Authors: Yuzi Luo, Huaji Qiu, Long Yin, Bo Liang, Fei Wang, Valery A Petrenko, Aihua LiuAbstract:Classical Swine Fever (CSF) is a devastating infectious disease caused by Classical Swine Fever virus (CSFV). The screening of CSFV-specific ligands is of great significance for diagnosis and treatment of CSF. Affinity selection from random peptide libraries is an efficient approach to discover ligands with high stability and specificity. Here, we screened phage ligands for the CSFV E2 protein from f8/8 landscape phage display library by biopanning and obtained four phage clones specific for the E2 protein of CSFV. Viral blocking assays indicated that the phage clone displaying the octapeptide sequence DRATSSNA remarkably inhibited the CSFV replication in PK-15 cells at a titer of 10(10) transduction units, as evidenced by significantly decreased viral RNA copies and viral titers. The phage-displayed E2-binding peptides have the potential to be developed as antivirals for CSF. (C) 2014 Elsevier B.V. All rights reserved.
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specific ligands for Classical Swine Fever virus screened from landscape phage display library
Antiviral Research, 2014Co-Authors: Yuzi Luo, Huaji Qiu, Long Yin, Bo Liang, Fei Wang, Valery A Petrenko, Min Du, Aihua LiuAbstract:Classical Swine Fever (CSF) is a devastating infectious disease caused by Classical Swine Fever virus (CSFV). The screening of CSFV-specific ligands is of great significance for diagnosis and treatment of CSF. Affinity selection from random peptide libraries is an efficient approach to discover ligands with high stability and specificity. Here, we screened phage ligands for the CSFV E2 protein from f8/8 landscape phage display library by biopanning and obtained four phage clones specific for the E2 protein of CSFV. Viral blocking assays indicated that the phage clone displaying the octapeptide sequence DRATSSNA remarkably inhibited the CSFV replication in PK-15 cells at a titer of 10(10) transduction units, as evidenced by significantly decreased viral RNA copies and viral titers. The phage-displayed E2-binding peptides have the potential to be developed as antivirals for CSF. (C) 2014 Elsevier B.V. All rights reserved.
Yuzi Luo - One of the best experts on this subject based on the ideXlab platform.
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isolation and characterization of a moderately virulent Classical Swine Fever virus emerging in china
Transboundary and Emerging Diseases, 2017Co-Authors: Yuzi Luo, Yuan Sun, Yonggang Liu, Jianlin Lei, Shuili Xia, Yimin Wang, Lina Shao, Xingyu Meng, Mo Zhou, Huaji QiuAbstract:Classical Swine Fever (CSF) is a devastating infectious disease of pigs caused by Classical Swine Fever virus (CSFV). In China, CSF has been under control owing to extensive vaccination with the lapinized attenuated vaccine (C-strain) since 1950s, despite sporadic or endemic in many regions. However, recently, CSF outbreaks occurred in a large number of Swine herds in China. Here, we isolated 15 CSFV strains from diverse C-strain-vaccinated pig farms in China and characterized the genetic variations and antigenicity of the new isolates. The new strains showed unique variations in the E2 protein and were clustered to the subgenotype 2.1d of CSFV recently emerging in China in the phylogenetic tree. Cross-neutralization test showed that the neutralizing titres of porcine anti-C-strain sera against the new isolates were substantially lower than those against both the highly virulent Shimen strain and the subgenotype 2.1b strains that were isolated in China in 2006 and 2009, respectively. In addition, experimental animal infection showed that the HLJZZ2014 strain-infected pigs displayed lower mortality and less severe clinical signs and pathological changes compared with the Shimen strain-infected pigs. The HLJZZ2014 strain was defined to be moderately virulent based on a previously established assessment system for CSFV virulence evaluation, and the virus shedding and the viral load in various tissues of the CSFV HLJZZ2014 strain-infected pigs were significantly lower than those of the Shimen strain-infected pigs. Taken together, the subgenotype 2.1d isolate of CSFV is a moderately virulent strain with molecular variations and antigenic alterations.
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interferon inducible oligoadenylate synthetase like protein acts as an antiviral effector against Classical Swine Fever virus via the mda5 mediated type i interferon signaling pathway
Journal of Virology, 2017Co-Authors: Yuexiu Zhang, Yuzi Luo, Yuan Sun, Jinghan Wang, Lingkai Zhang, Qian Yang, Huaji QiuAbstract:Classical Swine Fever virus (CSFV) is the causative agent of Classical Swine Fever (CSF), which poses a serious threat to the global pig industry. Interferons (IFNs) and IFN-stimulated genes (ISGs) play a key role in host antiviral defense. We have previously screened the porcine 2'-5'-oligoadenylate synthetase-like protein (pOASL) as a potential anti-CSFV ISG using a reporter CSFV. This study aimed to clarify the underlying antiviral mechanism of pOASL against CSFV. We confirmed that CSFV replication was significantly suppressed in lentivirus-delivered, pOASL-overexpressing PK-15 cells, whereas silencing the expression of endogenous pOASL by small interfering RNAs markedly enhanced CSFV growth. In addition, the transcriptional level of pOASL was upregulated both in vitro and in vivo upon CSFV infection. Interestingly, the anti-CSFV effects of pOASL are independent of the canonical RNase L pathway but depend on the activation of the type I IFN response. Glutathione S-transferase pulldown and coimmunoprecipitation assays revealed that pOASL interacts with MDA5, a double-stranded RNA sensor, and further enhances MDA5-mediated type I IFN signaling. Moreover, we showed that pOASL exerts anti-CSFV effects in an MDA5-dependent manner. In conclusion, pOASL suppresses CSFV replication via the MDA5-mediated type I IFN-signaling pathway.IMPORTANCE The host innate immune response plays an important role in mounting the initial resistance to viral infection. Here, we identify the porcine 2'-5'-oligoadenylate synthetase-like protein (pOASL) as an interferon (IFN)-stimulated gene (ISG) against Classical Swine Fever virus (CSFV). We demonstrate that the anti-CSFV effects of pOASL depend on the activation of type I IFN response. In addition, we show that pOASL, as an MDA5-interacting protein, is a coactivator of MDA5-mediated IFN induction to exert anti-CSFV actions. This work will be beneficial to the development of novel anti-CSFV strategies by targeting pOASL.
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guanylate binding protein 1 an interferon induced gtpase exerts an antiviral activity against Classical Swine Fever virus depending on its gtpase activity
Journal of Virology, 2016Co-Authors: Jinghan Wang, Xiao Wang, Yuzi Luo, Yuan Sun, Shuili Xia, Lingkai Zhang, Qian Yang, Yan Zhu, Muhammad Munir, Huaji QiuAbstract:UNLABELLED: Many viruses trigger the type I interferon (IFN) pathway upon infection, resulting in the transcription of hundreds of interferon-stimulated genes (ISGs), which define the antiviral state of the host. Classical Swine Fever virus (CSFV) is the causative agent of Classical Swine Fever (CSF), a highly contagious viral disease endangering the pig industry in many countries. However, anti-CSFV ISGs are poorly documented. Here we screened 20 ISGs that are commonly induced by type I IFNs against CSFV in lentivirus-delivered cell lines, resulting in the identification of guanylate-binding protein 1 (GBP1) as a potent anti-CSFV ISG. We observed that overexpression of GBP1, an IFN-induced GTPase, remarkably suppressed CSFV replication, whereas knockdown of endogenous GBP1 expression by small interfering RNAs significantly promoted CSFV growth. Furthermore, we demonstrated that GBP1 acted mainly on the early phase of CSFV replication and inhibited the translation efficiency of the internal ribosome entry site of CSFV. In addition, we found that GBP1 was upregulated at the transcriptional level in CSFV-infected PK-15 cells and in various organs of CSFV-infected pigs. Coimmunoprecipitation and glutathione S-transferase (GST) pulldown assays revealed that GBP1 interacted with the NS5A protein of CSFV, and this interaction was mapped in the N-terminal globular GTPase domain of GBP1. Interestingly, the K51 of GBP1, which is crucial for its GTPase activity, was essential for the inhibition of CSFV replication. We showed further that the NS5A-GBP1 interaction inhibited GTPase activity, which was critical for its antiviral effect. Taking our findings together, GBP1 is an anti-CSFV ISG whose action depends on its GTPase activity. IMPORTANCE: Classical Swine Fever virus (CSFV) is the causative agent of Classical Swine Fever (CSF), an economically important viral disease affecting the pig industry in many countries. To date, only a few host restriction factors against CSFV, including interferon-stimulated genes (ISGs), have been characterized. Using a minilibrary of porcine ISGs, we identify porcine guanylate-binding protein 1 (GBP1) as a potent antiviral ISG against CSFV. We further show that the anti-CSFV action of GBP1 depends on its GTPase activity. The K51 of GBP1, critical for its GTPase activity, is essential for the antiviral action of GBP1 against CSFV replication, and the binding of the NS5A protein to GBP1 antagonizes the GTPase activity and thus the antiviral effect. This study will facilitate the development of anti-CSFV therapeutic agents by targeting host factors and may provide a new strategy for the control of CSF.
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the laminin receptor is a cellular attachment receptor for Classical Swine Fever virus
Journal of Virology, 2015Co-Authors: Jianing Chen, Liang Shen, Hong Dong, Xiao Wang, Yuzi Luo, Yuan Sun, Huaji QiuAbstract:UNLABELLED Classical Swine Fever virus (CSFV) is the causative agent of Classical Swine Fever (CSF), a highly contagious, economically important viral disease in many countries. The E(rns) and E2 envelope glycoproteins are responsible for the binding to and entry into the host cell by CSFV. To date, only one cellular receptor, heparan sulfate (HS), has been identified as being involved in CSFV attachment. HS is also present on the surface of various cells that are nonpermissive to CSFV. Hence, there must be another receptor(s) that has been unidentified to date. In this study, we used a set of small interfering RNAs (siRNAs) against a number of porcine cell membrane protein genes to screen cellular proteins involved in CSFV infection. This approach resulted in the identification of several proteins, and of these, the laminin receptor (LamR) has been demonstrated to be a cellular receptor for several viruses. Confocal analysis showed that LamR is colocalized with CSFV virions on the membrane, and a coimmunoprecipitation assay indicated that LamR interacts with the CSFV E(rns) protein. In inhibition assays, anti-LamR antibodies, soluble laminin, or LamR protein significantly inhibited CSFV infection in a dose-dependent manner. Transduction of PK-15 cells with a recombinant lentivirus expressing LamR yielded higher viral titers. Moreover, an attachment assay demonstrated that LamR functions during virus attachment. We also demonstrate that LamR acts as an alternative attachment receptor, especially in SK6 cells. These results indicate that LamR is a cellular attachment receptor for CSFV. IMPORTANCE Classical Swine Fever virus (CSFV) is the causative agent of Classical Swine Fever (CSF), an economically important viral disease affecting the pig industry in many countries. To date, only heparan sulfate (HS) has been identified to be an attachment receptor for CSFV. Here, using RNA interference screening with small interfering RNAs (siRNAs) against a number of porcine membrane protein genes, we identified the laminin receptor (LamR) to be another attachment receptor. We demonstrate the involvement of LamR together with HS in virus attachment, and we elucidate the relationship between LamR and HS. LamR also serves as an attachment receptor for many viral pathogens, including dengue virus, a fatal human flavivirus. The study will help to enhance our understanding of the life cycle of flaviviruses and the development of antiviral strategies for flaviviruses.
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specific ligands for Classical Swine Fever virus screened from landscape phage display library
Antiviral Research, 2014Co-Authors: Yuzi Luo, Huaji Qiu, Long Yin, Bo Liang, Fei Wang, Valery A Petrenko, Aihua LiuAbstract:Classical Swine Fever (CSF) is a devastating infectious disease caused by Classical Swine Fever virus (CSFV). The screening of CSFV-specific ligands is of great significance for diagnosis and treatment of CSF. Affinity selection from random peptide libraries is an efficient approach to discover ligands with high stability and specificity. Here, we screened phage ligands for the CSFV E2 protein from f8/8 landscape phage display library by biopanning and obtained four phage clones specific for the E2 protein of CSFV. Viral blocking assays indicated that the phage clone displaying the octapeptide sequence DRATSSNA remarkably inhibited the CSFV replication in PK-15 cells at a titer of 10(10) transduction units, as evidenced by significantly decreased viral RNA copies and viral titers. The phage-displayed E2-binding peptides have the potential to be developed as antivirals for CSF. (C) 2014 Elsevier B.V. All rights reserved.
Martin Beer - One of the best experts on this subject based on the ideXlab platform.
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Classical Swine Fever an updated review
Viruses, 2017Co-Authors: Sandra Blome, Christoph Staubach, Julia Henke, Jolene Carlson, Martin BeerAbstract:Classical Swine Fever (CSF) remains one of the most important transboundary viral diseases of Swine worldwide. The causative agent is CSF virus, a small, enveloped RNA virus of the genus Pestivirus. Based on partial sequences, three genotypes can be distinguished that do not, however, directly correlate with virulence. Depending on both virus and host factors, a wide range of clinical syndromes can be observed and thus, laboratory confirmation is mandatory. To this means, both direct and indirect methods are utilized with an increasing degree of commercialization. Both infections in domestic pigs and wild boar are of great relevance; and wild boars are a reservoir host transmitting the virus sporadically also to pig farms. Control strategies for epidemic outbreaks in free countries are mainly based on Classical intervention measures; i.e., quarantine and strict culling of affected herds. In these countries, vaccination is only an emergency option. However, live vaccines are used for controlling the disease in endemically infected regions in Asia, Eastern Europe, the Americas, and some African countries. Here, we will provide a concise, updated review on virus properties, clinical signs and pathology, epidemiology, pathogenesis and immune responses, diagnosis and vaccination possibilities.
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Classical Swine Fever vaccines state of the art
Veterinary Microbiology, 2017Co-Authors: Sandra Blome, Ilona Reimann, Claudia Mos, Patricia Konig, Martin BeerAbstract:Due to its impact on animal health and pig industry, Classical Swine Fever (CSF) is still one of the most important viral diseases of pigs. To control the disease, safe and highly efficacious live attenuated vaccines exist for decades. These vaccines have usually outstanding efficacy and safety but lack differentiability of infected from vaccinated animals (DIVA or marker strategy). In contrast, the first generation of E2 subunit marker vaccines shows constraints in efficacy, application, and production. To overcome these limitations, new generations of marker vaccines are developed. A wide range of approaches have been tried including recombinant vaccines, recombinant inactivated vaccines or subunit vaccines, vector vaccines, and DNA/RNA vaccines. During the last years, especially attenuated deletion vaccines or chimeric constructs have shown potential. At present, especially two new constructs have been intensively tested, the adenovirus-delivered, Semliki Forest virus replicon-vectored marker vaccine candidate “rAdV-SFV-E2” and the pestivirus chimera “CP7_E2alf”. The later was recently licensed by the European Medicines Agency. Under field conditions, all marker vaccines have to be accompanied by a potent test system. Particularly this point shows still weaknesses and it is important to embed vaccination in a well-established vaccination strategy and a suitable diagnostic workflow. In summary, conventional vaccines are a standard in terms of efficacy. However, only vaccines with DIVA will allow improved eradication strategies e.g. also under emergency vaccination conditions in free regions. To answer this demand, new generations of marker vaccines have been developed and add now to the tool box of CSF control.
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genetic variability and distribution of Classical Swine Fever virus
Animal Health Research Reviews, 2015Co-Authors: Martin Beer, Christoph Staubach, Katja V Goller, Sandra BlomeAbstract:Classical Swine Fever is a highly contagious disease that affects domestic and wild pigs worldwide. The causative agent of the disease is Classical Swine Fever virus (CSFV), which belongs to the genus Pestivirus within the family Flaviviridae. On the genome level, CSFV can be divided into three genotypes with three to four sub-genotypes. Those genotypes can be assigned to distinct geographical regions. Knowledge about CSFV diversity and distribution is important for the understanding of disease dynamics and evolution, and can thus help to design optimized control strategies. For this reason, the geographical pattern of CSFV diversity and distribution are outlined in the presented review. Moreover, current knowledge with regard to genetic virulence markers or determinants and the role of the quasispecies composition is discussed.
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molecular epidemiology of current Classical Swine Fever virus isolates of wild boar in germany
Journal of General Virology, 2010Co-Authors: Immanuel Leifer, Christoph Staubach, Sandra Blome, Bernd Hoffmann, Dirk Hoper, Thomas Bruun Rasmussen, Gunter Strebelow, Detlef Horethbontgen, Martin BeerAbstract:Classical Swine Fever (CSF) has caused significant economic losses in industrialized pig production, and is still present in some European countries. Recent CSF outbreaks in Europe were mainly associated with strains of genogroup 2 (subgroup 2.3). Although there are extensive datasets regarding 2.3 strains, there is very little information available on longer fragments or whole Classical Swine Fever virus (CSFV) genomes. Furthermore, there are no detailed analyses of the molecular epidemiology of CSFV wild boar isolates available. Nevertheless, complete genome sequences are supportive in phylogenetic analyses, especially in affected wild boar populations. Here, German CSFV strains of subgroup 2.3 were fully sequenced using two different approaches: (i) a universal panel of CSFV primers that were developed to amplify the complete genome in overlapping fragments for chain-terminator sequencing; and (ii) generation of a single full-length amplicon of the CSFV genome obtained by long-range RT-PCR for deep sequencing with next-generation sequencing technology. In total, five different strains of CSFV subgroup 2.3 were completely sequenced using these newly developed protocols. The approach was used to study virus spread and evolutionary history in German wild boar. For the first time, the results of our study clearly argue for the possibility of a long-term persistence of genotype 2.3 CSFV strains in affected regions at an almost undetectable level, even after long-term oral vaccination campaigns with intensive monitoring. Hence, regional persistence in wild boar populations has to be taken into account as an important factor in the continual outbreaks in affected areas.
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chimeric pestiviruses candidates for live attenuated Classical Swine Fever marker vaccines
Journal of General Virology, 2007Co-Authors: Franziska Wehrle, Sandra Renzullo, Anja Faust, Martin Beer, Volker Kaden, Martin A HofmannAbstract:The use of attenuated Classical Swine Fever virus (CSFV) strains as live vaccines is no longer allowed for the control of Classical Swine Fever in Europe, due to the inability to differentiate between infected and vaccinated animals (Differentiating Infected from Vaccinated Animals; DIVA), except as emergency vaccines or as bait vaccines for wild boars. Thus, the establishment of a DIVA vaccine(s) is of pivotal importance for the control of this infectious disease. In this study, recombinant versions of the live-attenuated vaccine strain CSFV Riems were generated by replacing parts of the E2 gene with the corresponding sequence of border disease virus strain Gifhorn. Three cDNA clones were constructed: pRiems-ABC-Gif, pRiems-A-Gif and pRiems-BC-Gif. Infectious particles were obtained from clones pRiems-ABC-Gif and pRiems-BC-Gif only, whereas transfected RNA from clone pRiems-A-Gif behaved like a replicon. Based on its ability to be differentiated in vitro from wild-type CSFV by mAbs, vRiems-ABC-Gif was assessed for immunogenicity and protection against challenge infection in pigs. Before challenge, no CSFV-specific anti-E2 antibodies could be detected with commercial E2-blocking ELISAs in vRiems-ABC-Gif-vaccinated animals, whereas vRiems-vaccinated pigs developed high titres of anti-E2 antibodies, confirming the marker properties of this vaccine candidate. After oral vaccination, only partial protection against challenge infection was observed in the vRiems-ABC-Gif vaccinees, whereas all intramuscularly vaccinated animals and all vRiems-vaccinated animals were fully protected. These experiments suggest that the strategy of exchanging specific antigenic epitopes among pestiviruses is a promising tool for the development of new CSFV marker vaccines.