The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform

Andrew A. Mercer - One of the best experts on this subject based on the ideXlab platform.

  • Molecular Genetic Analysis of Orf Virus: A PoxVirus That Has Adapted to Skin
    MDPI AG, 2015
    Co-Authors: Stephen B. Fleming, Lyn M Wise, Andrew A. Mercer
    Abstract:

    Orf Virus is the type species of the ParapoxVirus genus of the family Poxviridae. It induces acute pustular skin lesions in sheep and goats and is transmissible to humans. The genome is G+C rich, 138 kbp and encodes 132 genes. It shares many essential genes with vaccinia Virus that are required for survival but encodes a number of unique factors that allow it to replicate in the highly specific immune environment of skin. Phylogenetic analysis suggests that both viral interleukin-10 and vascular endothelial growth factor genes have been “captured” from their host during the evolution of the parapoxViruses. Genes such as a chemokine binding protein and a protein that binds granulocyte-macrophage colony-stimulating factor and interleukin-2 appear to have evolved from a common poxVirus ancestral gene while three parapoxVirus nuclear factor (NF)-κB signalling pathway inhibitors have no homology to other known NF-κB inhibitors. A homologue of an anaphase-promoting complex subunit that is believed to manipulate the cell cycle and enhance viral DNA synthesis appears to be a specific adaptation for viral-replication in keratinocytes. The review focuses on the unique genes of Orf Virus, discusses their evolutionary origins and their role in allowing viral-replication in the skin epidermis

  • Orf Virus il 10 accelerates wound healing while limiting inflammation and scarring
    Wound Repair and Regeneration, 2014
    Co-Authors: Lyn M Wise, Stephen B. Fleming, Gabriella S Stuart, Nicola C Real, Andrew A. Mercer
    Abstract:

    Interleukin (IL)-10 plays a critical role in controlling wound inflammation and scar formation. Orf Virus, a zoonotic parapoxVirus, induces proliferative skin lesions that resolve with minimal scarring. Orf Virus encodes a range of factors that subvert the host's response to infection, including a homolog of IL-10. This study investigated, using a murine full-thickness wound model, whether purified Orf Virus IL-10 (ovIL-10) can regulate skin repair and scarring. Repeat injections of ovIL-10 into wounded skin accelerated wound closure. Histological analyses of wound sections revealed that treatment with ovIL-10 accelerated wound reepithelialization, granulation tissue coverage of the wound bed, and improved wound revascularization. In addition, wounds treated with ovIL-10 showed a reduction in macrophage infiltration, myofibroblast differentiation, and wound contraction. Treatment of wounds with ovIL-10 also resulted in a reduction in visible scarring that was consistent with the extent of scar tissue formed. Quantitative polymerase chain reaction analysis confirmed that ovIL-10 reduced the expression of key mediators of inflammation and granulation tissue formation. These findings show that ovIL-10, like mammalian IL-10, limits inflammation and scar tissue formation and reveal a new role for both mammalian and viral IL-10 in mediating tissue repair.

  • therapeutic immunomodulation using a Virus the potential of inactivated Orf Virus
    European Journal of Clinical Microbiology & Infectious Diseases, 2013
    Co-Authors: Olaf Weber, Andrew A. Mercer, Percy A. Knolle, Astrid Friebe, Hans-dieter Volk
    Abstract:

    Viruses can manipulate the immune response against them by various strategies to influence immune cells, i.e. by over-activation leading to functional inactivation, bypassing antigen presentation or even suppression of effector functions. Little is known, however, about how these features of immune regulation and modulation could be used for therapeutic purposes. Reasons for this include the complexity of immune regulatory mechanisms under certain disease conditions and the risks that infections with Viruses pose to human beings. The Orf Virus (OrfV), a member of the ParapoxVirus genus of the poxVirus family, is known as a common pathogen in sheep and goats worldwide. The inactivated OrfV, however, has been used as a preventative as well as therapeutic immunomodulator in veterinary medicine in different species. Here, we review the key results obtained in pre-clinical studies or clinical studies in veterinary medicine to characterise the therapeutic potential of inactivated OrfV. Inactivated OrfV has strong effects on cytokine secretion in mice and human immune cells, leading to an auto-regulated loop of initial up-regulation of inflammatory and Th1-related cytokines, followed by Th2-related cytokines that attenuate immunopathology. The therapeutic potential of inactivated OrfV has been recognised in several difficult-to-treat disease areas, such as chronic viral diseases, liver fibrosis or various forms of cancer. Further research will be required in order to evaluate the full beneficial potential of inactivated OrfV for therapeutic immunomodulation.

  • the vascular endothelial growth factor vegf e encoded by Orf Virus regulates keratinocyte proliferation and migration and promotes epidermal regeneration
    Cellular Microbiology, 2012
    Co-Authors: Lyn M Wise, Stephen B. Fleming, Gabriella S Stuart, Nicola C Real, Marie K Inder, Andrew A. Mercer
    Abstract:

    Summary Vascular endothelial growth factor (VEGF)-A, a key regulator of cutaneous blood vessel formation, appears to have an additional role during wound healing, enhancing re-epithelialization. Orf Virus, a zoonotic parapoxVirus, induces proliferative skin lesions that initiate in wounds and are characterized by extensive blood vessel formation, epidermal hyperplasia and rete ridge formation. The vascular changes beneath the lesion are largely due to viral-expressed VEGF-E. This study investigated using mouse skin models whether VEGF-E can induce epidermal changes such as that seen in the viral lesion. Injection of VEGF-E into normal skin increased the number of endothelial cells and blood vessels within the dermis and increased epidermal thickening and keratinocyte number. Injection of VEGF-E into wounded skin, which more closely mimics Orf Virus lesions, increased neo-epidermal thickness and area, promoted rete ridge formation, and enhanced wound re-epithelialization. Quantitative RT-PCR analysis showed that VEGF-E did not induce expression of epidermal-specific growth factors within the wound, but did increase matrix metalloproteinase (MMP)-2 and MMP-9 expression. In cell-based assays, VEGF-E induced keratinocyte migration and proliferation, responses that were inhibited by a neutralizing antibody against VEGF receptor (VEGFR)-2. These findings demonstrate that VEGF-E, both directly and indirectly, regulates keratinocyte function, thereby promoting epidermal regeneration.

  • Characterization of immunostimulatory components of Orf Virus (parapoxVirus ovis)
    Journal of General Virology, 2011
    Co-Authors: Astrid Friebe, Andrew A. Mercer, Angela Siegling, Hans-dieter Volk, Sonja Friederichs, Kai Scholz, Uwe Janssen, Corinna Scholz, Tobias Schlapp, Olaf Weber
    Abstract:

    Inactivated Orf Virus (OrfV, parapoxVirus ovis) induces antiviral activity in animal models of acute and chronic viral infections and exerts strong effects on human immune cells. OrfV activates antigen presenting cells (APC) via CD14 and, probably, Toll-like receptor signalling, and triggers the release of IFN-γ that has been identified as the key mediator of the antiviral activity. After delineating Virus proteins as being the most likely active constituent, we aimed to characterize the OrfV proteins responsible for the therapeutic effect. By using a vaccinia Virus/OrfV expression library we identified several multi-gene DNA fragments with strong immunomodulatory activity. Together these fragments contain 27 Orfs. The encoded proteins are related to virion structure and transcription but are otherwise unrelated. Two proteins were separately expressed and purified, and demonstrated immunostimulatory activity. Gene expression profiles induced by OrfV and the identified fragments were investigated by microarray analysis. Interestingly, all active fragments induced a similar gene-expression pattern, differing only in quantitative aspects. Obviously, several proteins of OrfV activate similar cellular pathways, modulating APC to generate a strong T-helper 1-dominated immune response. This was balanced by additional induction of immune dampening mechanisms, suggesting regulatory differences compared to single cytokine therapies. We conclude that OrfV may have the potential to enrich the armamentarium of antiviral therapies.

Stephen B. Fleming - One of the best experts on this subject based on the ideXlab platform.

  • Molecular Genetic Analysis of Orf Virus: A PoxVirus That Has Adapted to Skin
    MDPI AG, 2015
    Co-Authors: Stephen B. Fleming, Lyn M Wise, Andrew A. Mercer
    Abstract:

    Orf Virus is the type species of the ParapoxVirus genus of the family Poxviridae. It induces acute pustular skin lesions in sheep and goats and is transmissible to humans. The genome is G+C rich, 138 kbp and encodes 132 genes. It shares many essential genes with vaccinia Virus that are required for survival but encodes a number of unique factors that allow it to replicate in the highly specific immune environment of skin. Phylogenetic analysis suggests that both viral interleukin-10 and vascular endothelial growth factor genes have been “captured” from their host during the evolution of the parapoxViruses. Genes such as a chemokine binding protein and a protein that binds granulocyte-macrophage colony-stimulating factor and interleukin-2 appear to have evolved from a common poxVirus ancestral gene while three parapoxVirus nuclear factor (NF)-κB signalling pathway inhibitors have no homology to other known NF-κB inhibitors. A homologue of an anaphase-promoting complex subunit that is believed to manipulate the cell cycle and enhance viral DNA synthesis appears to be a specific adaptation for viral-replication in keratinocytes. The review focuses on the unique genes of Orf Virus, discusses their evolutionary origins and their role in allowing viral-replication in the skin epidermis

  • Orf Virus il 10 accelerates wound healing while limiting inflammation and scarring
    Wound Repair and Regeneration, 2014
    Co-Authors: Lyn M Wise, Stephen B. Fleming, Gabriella S Stuart, Nicola C Real, Andrew A. Mercer
    Abstract:

    Interleukin (IL)-10 plays a critical role in controlling wound inflammation and scar formation. Orf Virus, a zoonotic parapoxVirus, induces proliferative skin lesions that resolve with minimal scarring. Orf Virus encodes a range of factors that subvert the host's response to infection, including a homolog of IL-10. This study investigated, using a murine full-thickness wound model, whether purified Orf Virus IL-10 (ovIL-10) can regulate skin repair and scarring. Repeat injections of ovIL-10 into wounded skin accelerated wound closure. Histological analyses of wound sections revealed that treatment with ovIL-10 accelerated wound reepithelialization, granulation tissue coverage of the wound bed, and improved wound revascularization. In addition, wounds treated with ovIL-10 showed a reduction in macrophage infiltration, myofibroblast differentiation, and wound contraction. Treatment of wounds with ovIL-10 also resulted in a reduction in visible scarring that was consistent with the extent of scar tissue formed. Quantitative polymerase chain reaction analysis confirmed that ovIL-10 reduced the expression of key mediators of inflammation and granulation tissue formation. These findings show that ovIL-10, like mammalian IL-10, limits inflammation and scar tissue formation and reveal a new role for both mammalian and viral IL-10 in mediating tissue repair.

  • the vascular endothelial growth factor vegf e encoded by Orf Virus regulates keratinocyte proliferation and migration and promotes epidermal regeneration
    Cellular Microbiology, 2012
    Co-Authors: Lyn M Wise, Stephen B. Fleming, Gabriella S Stuart, Nicola C Real, Marie K Inder, Andrew A. Mercer
    Abstract:

    Summary Vascular endothelial growth factor (VEGF)-A, a key regulator of cutaneous blood vessel formation, appears to have an additional role during wound healing, enhancing re-epithelialization. Orf Virus, a zoonotic parapoxVirus, induces proliferative skin lesions that initiate in wounds and are characterized by extensive blood vessel formation, epidermal hyperplasia and rete ridge formation. The vascular changes beneath the lesion are largely due to viral-expressed VEGF-E. This study investigated using mouse skin models whether VEGF-E can induce epidermal changes such as that seen in the viral lesion. Injection of VEGF-E into normal skin increased the number of endothelial cells and blood vessels within the dermis and increased epidermal thickening and keratinocyte number. Injection of VEGF-E into wounded skin, which more closely mimics Orf Virus lesions, increased neo-epidermal thickness and area, promoted rete ridge formation, and enhanced wound re-epithelialization. Quantitative RT-PCR analysis showed that VEGF-E did not induce expression of epidermal-specific growth factors within the wound, but did increase matrix metalloproteinase (MMP)-2 and MMP-9 expression. In cell-based assays, VEGF-E induced keratinocyte migration and proliferation, responses that were inhibited by a neutralizing antibody against VEGF receptor (VEGFR)-2. These findings demonstrate that VEGF-E, both directly and indirectly, regulates keratinocyte function, thereby promoting epidermal regeneration.

  • A Novel Bcl-2-Like Inhibitor of Apoptosis Is Encoded by the ParapoxVirus Orf Virus
    Journal of virology, 2007
    Co-Authors: Dana Westphal, Stephen B. Fleming, Elizabeth C. Ledgerwood, Merilyn Hibma, Ellena M. Whelan, Andrew A. Mercer
    Abstract:

    Apoptotic cell death forms part of the host defense against Virus infection. We tested Orf Virus, a member of the poxVirus family, for the ability to inhibit apoptosis and found that Orf Virus-infected cells were fully resistant to UV-induced changes in cell morphology, caspase activation, and DNA fragmentation. By using a library of vaccinia Virus-Orf Virus recombinants, we identified an Orf Virus gene (OrfV125) whose presence was linked with the inhibition of apoptosis. The 173-amino-acid predicted protein had no clear homologs in public databases other than those encoded by other parapoxViruses. However, OrfV125 possessed a distinctive C-terminal domain which was necessary and sufficient to direct the protein to the mitochondria. We determined that OrfV125 alone could fully inhibit UV-induced DNA fragmentation, caspase activation, and cytochrome c release and that its mitochondrial localization was required for its antiapoptotic function. In contrast, OrfV125 did not prevent UV-induced activation of c-Jun NH2-terminal kinase, an event occurring upstream of the mitochondria. These features are comparable to the antiapoptotic properties of the mitochondrial regulator Bcl-2. Furthermore, bioinformatic analyses revealed sequence and secondary-structure similarities to Bcl-2 family members, including characteristic residues of all four Bcl-2 homology domains. Consistent with this, the viral protein inhibited the UV-induced activation of the proapoptotic Bcl-2 family members Bax and Bak. OrfV125 is the first parapoxVirus apoptosis inhibitor to be identified, and we propose that it is a new antiapoptotic member of the Bcl-2 family.

  • comparative analysis of genome sequences of three isolates of Orf Virus reveals unexpected sequence variation
    Virus Research, 2006
    Co-Authors: Andrew A. Mercer, Kate M Fraser, Norihito Ueda, Sonjamaria Friederichs, Kay Hofmann, Trudie Bateman, Stephen B. Fleming
    Abstract:

    Abstract Orf Virus (OrfV) is the type species of the ParapoxVirus genus. Here, we present the genomic sequence of the most well studied OrfV isolate, strain NZ2. The NZ2 genome is 138 kbp and contains 132 putative genes, 88 of which are present in all analyzed chordopoxViruses. Comparison of the NZ2 genome with the genomes of 2 other fully sequenced isolates of OrfV revealed that all 3 genomes carry each of the 132 genes, but there are substantial sequence variations between isolates in a significant number of genes, including 9 with inter-isolate amino acid sequence identity of only 38–79%. Each genome has an average of 64% G + C but each has a distinctive pattern of substantial deviation from the average within particular regions of the genome. The same pattern of variation was also seen in the genome of another parapoxVirus species and was clearly unlike the uniform patterns of G + C content seen in all other genera of chordopoxViruses. The availability of genomic sequences of three Orf Virus isolates allowed us to more accurately assess likely coding regions and thereby revise published data for 24 genes and to predict two previously unrecognized genes.

Lyn M Wise - One of the best experts on this subject based on the ideXlab platform.

  • Molecular Genetic Analysis of Orf Virus: A PoxVirus That Has Adapted to Skin
    MDPI AG, 2015
    Co-Authors: Stephen B. Fleming, Lyn M Wise, Andrew A. Mercer
    Abstract:

    Orf Virus is the type species of the ParapoxVirus genus of the family Poxviridae. It induces acute pustular skin lesions in sheep and goats and is transmissible to humans. The genome is G+C rich, 138 kbp and encodes 132 genes. It shares many essential genes with vaccinia Virus that are required for survival but encodes a number of unique factors that allow it to replicate in the highly specific immune environment of skin. Phylogenetic analysis suggests that both viral interleukin-10 and vascular endothelial growth factor genes have been “captured” from their host during the evolution of the parapoxViruses. Genes such as a chemokine binding protein and a protein that binds granulocyte-macrophage colony-stimulating factor and interleukin-2 appear to have evolved from a common poxVirus ancestral gene while three parapoxVirus nuclear factor (NF)-κB signalling pathway inhibitors have no homology to other known NF-κB inhibitors. A homologue of an anaphase-promoting complex subunit that is believed to manipulate the cell cycle and enhance viral DNA synthesis appears to be a specific adaptation for viral-replication in keratinocytes. The review focuses on the unique genes of Orf Virus, discusses their evolutionary origins and their role in allowing viral-replication in the skin epidermis

  • Orf Virus il 10 accelerates wound healing while limiting inflammation and scarring
    Wound Repair and Regeneration, 2014
    Co-Authors: Lyn M Wise, Stephen B. Fleming, Gabriella S Stuart, Nicola C Real, Andrew A. Mercer
    Abstract:

    Interleukin (IL)-10 plays a critical role in controlling wound inflammation and scar formation. Orf Virus, a zoonotic parapoxVirus, induces proliferative skin lesions that resolve with minimal scarring. Orf Virus encodes a range of factors that subvert the host's response to infection, including a homolog of IL-10. This study investigated, using a murine full-thickness wound model, whether purified Orf Virus IL-10 (ovIL-10) can regulate skin repair and scarring. Repeat injections of ovIL-10 into wounded skin accelerated wound closure. Histological analyses of wound sections revealed that treatment with ovIL-10 accelerated wound reepithelialization, granulation tissue coverage of the wound bed, and improved wound revascularization. In addition, wounds treated with ovIL-10 showed a reduction in macrophage infiltration, myofibroblast differentiation, and wound contraction. Treatment of wounds with ovIL-10 also resulted in a reduction in visible scarring that was consistent with the extent of scar tissue formed. Quantitative polymerase chain reaction analysis confirmed that ovIL-10 reduced the expression of key mediators of inflammation and granulation tissue formation. These findings show that ovIL-10, like mammalian IL-10, limits inflammation and scar tissue formation and reveal a new role for both mammalian and viral IL-10 in mediating tissue repair.

  • the vascular endothelial growth factor vegf e encoded by Orf Virus regulates keratinocyte proliferation and migration and promotes epidermal regeneration
    Cellular Microbiology, 2012
    Co-Authors: Lyn M Wise, Stephen B. Fleming, Gabriella S Stuart, Nicola C Real, Marie K Inder, Andrew A. Mercer
    Abstract:

    Summary Vascular endothelial growth factor (VEGF)-A, a key regulator of cutaneous blood vessel formation, appears to have an additional role during wound healing, enhancing re-epithelialization. Orf Virus, a zoonotic parapoxVirus, induces proliferative skin lesions that initiate in wounds and are characterized by extensive blood vessel formation, epidermal hyperplasia and rete ridge formation. The vascular changes beneath the lesion are largely due to viral-expressed VEGF-E. This study investigated using mouse skin models whether VEGF-E can induce epidermal changes such as that seen in the viral lesion. Injection of VEGF-E into normal skin increased the number of endothelial cells and blood vessels within the dermis and increased epidermal thickening and keratinocyte number. Injection of VEGF-E into wounded skin, which more closely mimics Orf Virus lesions, increased neo-epidermal thickness and area, promoted rete ridge formation, and enhanced wound re-epithelialization. Quantitative RT-PCR analysis showed that VEGF-E did not induce expression of epidermal-specific growth factors within the wound, but did increase matrix metalloproteinase (MMP)-2 and MMP-9 expression. In cell-based assays, VEGF-E induced keratinocyte migration and proliferation, responses that were inhibited by a neutralizing antibody against VEGF receptor (VEGFR)-2. These findings demonstrate that VEGF-E, both directly and indirectly, regulates keratinocyte function, thereby promoting epidermal regeneration.

  • vascular endothelial growth factors encoded by Orf Virus show surprising sequence variation but have a conserved functionally relevant structure
    Journal of General Virology, 2002
    Co-Authors: Andrew A. Mercer, Stephen B. Fleming, Catherine A. Mccaughan, Lyn M Wise, Colin Mcinnes, Alessandra Scagliarini, Mathias Buttner, Norihito Ueda, Hannsjoachim Rziha, P F Nettleton
    Abstract:

    The first report of a vascular endothelial growth factor (VEGF)-like gene in Orf Virus included the surprising observation that the genes from two isolates (NZ2 and NZ7) shared only 41·1% amino acid sequence identity. We have examined this sequence disparity by determining the VEGF gene sequence of 21 isolates of Orf Virus derived from diverse sources. Most isolates carried NZ2-like VEGF genes but their predicted amino acid sequences varied by up to 30·8% with an average amino acid identity between pairs of NZ2-like sequences of 86·1%. This high rate of sequence variation is more similar to interspecies than intraspecies variability. In contrast, only three isolates carried an NZ7-like VEGF gene and these varied from the NZ7 sequence by no more than a single nucleotide. The VEGF family are ligands for a set of tyrosine kinase receptors. The viral VEGFs are unique among the family in that they recognize VEGF receptor 2 (VEGFR-2) but not VEGFR-1 or VEGFR-3. Comparisons of the viral VEGFs with other family members revealed some correlations between conserved residues and the ability to recognize specific VEGF receptors. Despite the sequence variations, structural predictions for the viral VEGFs were very similar to each other and to the structure determined by X-ray crystallography for human VEGF-A. Structural modelling also revealed that a groove seen in the VEGF-A homodimer and believed to play a role in its binding to VEGFR-1 is blocked in the viral VEGFs. This may contribute to the inability of the viral VEGFs to bind VEGFR-1.

  • vascular endothelial growth factor vegf like protein from Orf Virus nz2 binds to vegfr2 and neuropilin 1
    Proceedings of the National Academy of Sciences of the United States of America, 1999
    Co-Authors: Lyn M Wise, Stephen B. Fleming, Andrew A. Mercer, Loreen J Savory, Tanja Veikkola, Carol Caesar, Angela A Vitali, Taija Makinen, Kari Alitalo, Steven A Stacker
    Abstract:

    Orf Virus, a member of the poxVirus family, produces a pustular dermatitis in sheep, goats, and humans. The lesions induced after infection with Orf Virus show extensive proliferation of vascular endothelial cells, dilation of blood vessels and dermal swelling. An explanation for the nature of these lesions may lie in the discovery that Orf Virus encodes an apparent homolog of the mammalian vascular endothelial growth factor (VEGF) family of molecules. These molecules mediate endothelial cell proliferation, vascular permeability, angiogenesis, and lymphangiogenesis via the endothelial cell receptors VEGFR-1 (Flt1), VEGFR-2 (KDR/Flk1), and VEGFR-3 (Flt4). The VEGF-like protein of Orf Virus strain NZ2 (OrfV2-VEGF) is most closely related in primary structure to VEGF. In this study we examined the biological activities and receptor specificity of the OrfV2-VEGF protein. OrfV2-VEGF was found to be a disulfide-linked homodimer with a subunit of ≈25 kDa. OrfV2-VEGF showed mitogenic activity on bovine aortic and human microvascular endothelial cells and induced vascular permeability. OrfV2-VEGF was found to bind and induce autophosphorylation of VEGFR-2 and was unable to bind or activate VEGFR-1 and VEGFR-3, but bound the newly identified VEGF165 receptor neuropilin-1. These results indicate that, from a functional viewpoint, OrfV2-VEGF is indeed a member of the VEGF family of molecules, but is unique, however, in that it utilizes only VEGFR-2 and neuropilin-1.

Shuhong Luo - One of the best experts on this subject based on the ideXlab platform.

  • identification of host cellular proteins lage3 and igfbp6 that interact with Orf Virus protein Orfv024
    Gene, 2018
    Co-Authors: Mingjian Long, Shuhong Luo, Wenbo Hao, Daxiang Chen, D L Rock, Daoyuan Gong, Yong Wang, Yuanyuan Wang, Ruixue Wang
    Abstract:

    Abstract Objective Orf Virus (OrfV) is the pathogen causing contagious pustular dermatitis in goats, sheep and herdsmen. Evidence has confirmed that OrfV can be used as a preventive and therapeutic immunomodulatory agent in several animal models. Our previous data demonstrated that OrfV024 is able to inhibit activation of the NF-κB signaling pathway and act as an important modulator for early immune responses against viral infection. However, the molecular mechanism by which OrfV024 exerting biological function remains unclear. In the present study, we explored and analyzed the function of host cellular proteins that interact with OrfV024. Methods The yeast two-hybrid (Y2H) assay was performed to screen proteins interacting with OrfV024 using a cDNA library derived from primary ovine fetal turbinate cells (OFTu). Two of the screened proteins were further confirmed by confocal microscopy, His-tag pull-down assay and CO-Immunoprecipitation (CO-IP) assay. In addition, the OrfV024 interaction network was constructed using the STRING database. Results In this study, 11 ovine cellular proteins were found to interact with OrfV024. In view of the importance of LAGE3 and IGFBP6 in the OrfV024 functional analysis, we further constructed LAGE3 and IGFBP6 interaction networks. The interactions between OrfV024 and LAGE3 or IGFBP6 were confirmed by confocal microscopy, LAGE3 was further confirmed in the His-tag pull-down assay and CO-IP assay. Conclusions Our findings indicate that OrfV024 can interact with ovine cellular proteins LAGE3 and IGFBP6.

  • Orf Virus 002 protein targets ovine protein s100a4 and inhibits nf κb signaling
    Frontiers in Microbiology, 2016
    Co-Authors: Daxiang Chen, Wenbo Hao, Mingjian Long, Bin Xiao, Huiqin Chen, D L Rock, Zewei Zheng, Shuhong Luo
    Abstract:

    Orf Virus (OrfV), a member of ParapoxVirus, has evolved various strategies to modulate the immune responses of host cells. The OrfV-encoded protein OrfV002, a regulator factor, has been found to inhibit the acetylation of NF-κB-p65 by blocking phosphorylation of NF-kB-p65 at Ser276 and also to disrupt the binding of NF-kB-p65 and p300. To explore the mechanism by which OrfV002 regulates NF-κB signaling, the understanding of OrfV002 potential binding partners in host cells is critical. In this study, ovine S100 calcium binding protein A4 (S100A4), prolylendopeptidase-like (PREPL) and NADH dehydrogenase (ubiquinone) 1 alpha subcomplex 8 (NDUFA8) were found to interact with OrfV002 based on the yeast two-hybrid (Y2H) assay using a cDNA library derived from primary ovine fetal turbinate cells (OFTu). GST pull-down and bidirectional co-immunoprecipitation assay results demonstrate that OrfV002 interacts with S100A4 directly. Following the pEGFP-OrfV002 (p002GFP) transfection, we found that cytoplasmic S100A4 translocates into the nucleus and co-localizes with OrfV002. Furthermore, the inhibitory effect of OrfV002 on NF-κB signaling was significantly restored by S100A4 knock-down phenotype, suggesting ovine S100A4 participating in the OrfV002-mediated NF-κB signaling. These data demonstrate that OrfV002 inhibits the NF-κB activation through its interaction with S100A4 along with its nucleus translocation.

  • Orf Virus 002 protein targets ovine protein S100A4 and inhibits NF-kappa B signaling
    Frontiers Media S.A., 2016
    Co-Authors: Daxiang Chen, Daniel L. Rock, Wenbo Hao, Mingjian Long, Bin Xiao, Huiqin Chen, Zewei Zheng, Shuhong Luo
    Abstract:

    Orf Virus (OrfV), a member of ParapoxVirus, has evolved various strategies to modulate the immune responses of host cells. The OrfV-encoded protein OrfV002, a regulator factor, has been found to inhibit the acetylation of NF-κB-p65 by blocking phosphorylation of NF-kB-p65 at Ser276 and also to disrupt the binding of NF-kB-p65 and p300. To explore the mechanism by which OrfV002 regulates NF-κB signaling, the understanding of OrfV002 potential binding partners in host cells is critical. In this study, ovine S100 calcium binding protein A4 (S100A4), prolylendopeptidase-like (PREPL) and NADH dehydrogenase (ubiquinone) 1 alpha subcomplex 8 (NDUFA8) were found to interact with OrfV002 based on the yeast two-hybrid (Y2H) assay using a cDNA library derived from primary ovine fetal turbinate cells (OFTu). GST pull-down and bidirectional co-immunoprecipitation assay results demonstrate that OrfV002 interacts with S100A4 directly. Following the pEGFP-OrfV002 (p002GFP) transfection, we found that cytoplasmic S100A4 translocates into the nucleus and co-localizes with OrfV002. Furthermore, the inhibitory effect of OrfV002 on NF-κB signaling was significantly restored by S100A4 knock-down phenotype, suggesting ovine S100A4 participating in the OrfV002-mediated NF-κB signaling. These data demonstrate that OrfV002 inhibits the NF-κB activation through its interaction with S100A4 along with its nucleus translocation

  • comparative genomic sequence analysis of chinese Orf Virus strain na1 11 with other parapoxViruses
    Archives of Virology, 2015
    Co-Authors: Wenbo Hao, D L Rock, Deguang Song, Feng Gao, Yongzheng Peng, Chaohui Duan, Chengbi Tong, Shuhong Luo
    Abstract:

    Orf Virus (OrfV) is a typical member of the genus ParapoxVirus. The parapoxVirus genome consists of highly variable terminal regions and relatively conserved central regions with a high G + C content. In our previous study, a novel OrfV strain, NA1/11, was isolated from northeastern China. To fully characterize this strain, we sequenced the entire genome of NA1/11 and conducted a comparative analysis using multiple parapoxViruses. The genomic sequence of NA1/11 was found to consist of 137,080 nucleotides with a G + C content of 63.6 %, but it did not contain the terminal hairpin sequence. Alignment of Orfs from NA1/11 with NZ2, IA82 and SA00 revealed several highly variable Orfs, while the most evident ones are Orfs 001, 103, 109–110, 116 and 132. An odd phenomenon in the region of Orfs 118–120 is that the non-coding fragments are almost as long as the coding fragments. By comparative analysis of inverted terminal repeats, we identified one repeat motif and a long conserved fragment. By comparing the ITRs of SA00 with those of three other OrfVs, more clues were obtained about the correlation between ITR sequence and host adaption. Comparison of the NA1/11 genome with the sequences of other strains of OrfV revealed highly variable regions, thus providing new insights into the genetic diversity of OrfV.

  • heterogeneity among Orf Virus isolates from goats in fujian province southern china
    PLOS ONE, 2013
    Co-Authors: Xuelin Chi, Shuhong Luo, Wenbo Hao, Xiancheng Zeng, Xiaohong Huang, Shihua Wang
    Abstract:

    Orf Virus is a parapoxVirus that causes recurring contagious ecthyma or Orf disease in goat, sheep and other wild and domestic ruminants. Infected animals show signs of pustular lesions on the mouth and muzzle and develop scabs over the lesions. Although the infection is usually cleared within 1–2 months, delayed growth and associated secondary infections could still impact the herds. Orf Virus can also infect humans, causing lesions similar to the animals in pathological histology. Prior infection of Orf Virus apparently offers little protective immunity against future infections. Several gene products of Orf Virus have been identified as responsible for immunomodulatory functions. In our recent study of Orf Virus isolates from an area along the Minjiang River in northern Fujian Province, we found a high heterogeneity among isolates from 10 farms within a 120-kilometer distance. Only two isolates from locations within 1 km to each other had same viral genes. There is no correlation between the geographical distance between the corresponding collection sites and the phylogenetic distance in OrfV011 or ORV059 genes for any two isolates. This finding suggests that there are diverse populations of Orf Virus present in the environment. This may in part contribute to the phenomenon of recurring outbreaks and heighten the need for better surveillance.

Hannsjoachim Rziha - One of the best experts on this subject based on the ideXlab platform.

  • Genomic Characterization of Orf Virus Strain D1701-V (ParapoxVirus) and Development of Novel Sites for Multiple Transgene Expression
    MDPI AG, 2019
    Co-Authors: Hannsjoachim Rziha, Mathias Buttner, Melanie Müller, Ferdinand Salomon, Alena Reguzova, Dominic Laible, Ralf Amann
    Abstract:

    The Orf Virus (OrfV; ParapoxVirus) strain D1701 with an attenuated phenotype and excellent immunogenic capacity is successfully used for the generation of recombinant vaccines against different viral infections. Adaption for growth in Vero cells was accompanied by additional major genomic changes resulting in OrfV strain variant D1701-V. In this study, restriction enzyme mapping, blot hybridization and DNA sequencing of the deleted region s (A, AT and D) in comparison to the predecessor strain D1701-B revealed the loss of 7 open reading frames (Orf008, Orf101, Orf102, Orf114, Orf115, Orf116, Orf117). The suitability of deletion site D for expression of foreign genes is demonstrated using novel synthetic early promoter eP1 and eP2. Comparison of promoter strength showed that the original vegf-e promoter Pv as well as promoter eP2 display an up to 11-fold stronger expression than promoter eP1, irrespective of the insertion site. Successful integration and expression of the fluorescent marker genes is demonstrated by gene- and insertion-site specific PCR assays, fluorescence microscopy and flow cytometry. For the first time OrfV recombinants are generated simultaneously expressing transgenes in two different insertion loci. That allows production of polyvalent vaccines containing several antigens against one or different pathogens in a single vectored OrfV vaccine

  • a new rabies vaccine based on a recombinant Orf Virus parapoxVirus expressing the rabies Virus glycoprotein
    Journal of Virology, 2013
    Co-Authors: Ralf Amann, Jorg Rohde, Ulrich Wulle, Douglas Conlee, Rudiger Raue, Olivier Martinon, Hannsjoachim Rziha
    Abstract:

    The present study describes the generation of a new Orf Virus (OrfV) recombinant, D1701-V-RabG, expressing the rabies Virus (RABV) glycoprotein that is correctly presented on the surface of infected cells without the need of replication or production of infectious recombinant Virus. One single immunization with recombinant OrfV can stimulate high RABV-specific Virus-neutralizing antibody (VNA) titers in mice, cats, and dogs, representing all nonpermissive hosts for the OrfV vector. The protective immune response against severe lethal challenge infection was analyzed in detail in mice using different dosages, numbers, and routes for immunization with the OrfV recombinant. Long-term levels of VNA could be elicited that remained greater than 0.5 IU per ml serum, indicative for the protective status. Single applications of higher doses (10(7) PFU) can be sufficient to confer complete protection against intracranial (i.c.) challenge, whereas booster immunization was needed for protection by the application of lower dosages. Anamnestic immune responses were achieved by each of the seven tested routes of inoculation, including oral application. Finally, in vivo antibody-mediated depletion of CD4-positive and/or CD8-posititve T cell subpopulations during immunization and/or challenge infection attested the importance of CD4 T cells for the induction of protective immunity by D1701-V-RabG. This report demonstrates another example of the potential of the OrfV vector and also indicates the capability of the new recombinant for vaccination of animals.

  • Orf Virus interferes with mhc class i surface expression by targeting vesicular transport and golgi
    BMC Veterinary Research, 2012
    Co-Authors: Hannsjoachim Rziha, Jorg Rohde, Frederic Emschermann, Michael R Knittler
    Abstract:

    Background: The Orf Virus (OrfV), a zoonotic ParapoxVirus, causes pustular skin lesions in small ruminants (goat and sheep). Intriguingly, OrfV can repeatedly infect its host, despite the induction of a specific immunity. These immune modulating and immune evading properties are still unexplained. Results: Here, we describe that OrfV infection of permissive cells impairs the intracellular transport of MHC class I molecules (MHC I) as a result of structural disruption and fragmentation of the Golgi apparatus. Depending on the duration of infection, we observed a pronounced co-localization of MHC I and COP-I vesicular structures as well as a reduction of MHC I surface expression of up to 50%. These subversion processes are associated with early OrfV gene expression and are accompanied by disturbed carbohydrate trimming of post-ER MHC I. The MHC I population remaining on the cell surface shows an extended half-life, an effect that might be partially controlled also by late OrfV genes. Conclusions: The presented data demonstrate that OrfV down-regulates MHC I surface expression in infected cells by targeting the late vesicular export machinery and the structure and function of the Golgi apparatus, which might aid to escape cellular immune recognition.

  • a new recombinant Orf Virus Orfv parapoxVirus protects rabbits against lethal infection with rabbit hemorrhagic disease Virus rhdv
    Vaccine, 2011
    Co-Authors: Joerg Rohde, Horst Schirrmeier, Harald Granzow, Hannsjoachim Rziha
    Abstract:

    This report describes the generation of a new recombinant Orf Virus (OrfV; ParapoxVirus) expressing the major capsid protein VP1 (VP60) of the caliciVirus, rabbit hemorrhagic disease Virus (RHDV). Authentic expression of VP1 could be demonstrated in cells infected with the recombinant D1701-V-VP1 without the need for production of infectious OrfV progeny. Notably, infected cells also released empty caliciVirus-like particles (VLPs). Challenge experiments showed that even a single immunization with =105 PFU of D1701-V-VP1 protected rabbits against lethal RHDV infection. ELISA tests indicated that the protective immunity mediated by D1701-V-VP1 did not strictly depend on the presence of detectable RHDV-specific serum antibodies. The induction of interleukin-2 found only in the sera of rabbits immunized with the D1701-V-VP1, but not in sera of rabbits immunized with the inactivated commercial vaccine RIKA-VACC, might indicate also some involvement of T-cells in protection. Collectively, this work adds another example of the successful use of the OrfV vector system for the generation of a recombinant vaccine, and demonstrates its potential as an alternative vaccine to protect rabbits against RHDV infection.

  • prime boost immunization using dna vaccine and recombinant Orf Virus protects pigs against pseudorabies Virus herpes suid 1
    Vaccine, 2006
    Co-Authors: Daniel Dory, Hannsjoachim Rziha, Timo Fischer, Veronique Beven, Roland Cariolet, Andre Jestin
    Abstract:

    Abstract The present study demonstrates the protective potential of a novel prime-boost vaccination strategy of pigs against lethal Pseudorabies Virus (PRV; Herpes suid 1) infection. Animals were primed with Sindbis Virus-derived plasmids that express viral glycoproteins gC and gD (gC- and gD-pSIN) and subsequently booster immunized with Orf Virus (OrfV; ParapoxVirus ) recombinants expressing gC and gD (D1701-VrVgC and -VrVgD). The prime-boost vaccination induced strong humoral and cellular-like PRV-specific immune responses. All prime-boost vaccinated pigs survived the lethal challenge infection without PRV-specific clinical symptoms and presented excellent body weight loss attenuation. Most notably, nasal shedding of challenge Virus was reduced by more than about 3 log 10 , clearly reducing the risk of infection of non-immunized pigs.