The Experts below are selected from a list of 273 Experts worldwide ranked by ideXlab platform
Inger K. Damon - One of the best experts on this subject based on the ideXlab platform.
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use of live Variola Virus to determine whether cast eij mice are a suitable surrogate animal model for human smallpox
Virus Research, 2020Co-Authors: Nadia F Gallardoromero, Victoria A. Olson, Scott K. Smith, Ashley V. Kondas, Darin S. Carroll, Christina L. Hutson, Johanna S. Salzer, Paul Hudson, Sharon Dietzostergaard, Inger K. DamonAbstract:Numerous animal models of systemic orthopoxVirus disease have been developed to evaluate therapeutics against Variola Virus (VARV), the causative agent of smallpox. These animal models do not resemble the disease presentation in human smallpox and most used surrogate OrthopoxViruses. A rodent model using VARV has a multitude of advantages, and previous investigations identified the CAST/EiJ mouse as highly susceptible to monkeypox Virus infection, making it of interest to determine if these rodents are also susceptible to VARV infection. In this study, we inoculated CAST/EiJ mice with a range of VARV doses (102-106 plaque forming units). Some animals had detectable viable VARV from the oropharynx between days 3 and 12 post inoculation. Despite evidence of disease, the CAST/EiJ mouse does not provide a model for clinical smallpox due to mild signs of morbidity and limited skin lesions. However, in contrast to previous rodent models using VARV challenge (i.e. prairie dogs and SCID mice), a robust immune response was observed in the CAST/EiJ mice (measured by Immunoglobulin G enzyme-linked immunosorbent assay). This is an advantage of this model for the study of VARV and presents a unique potential for the study of the immunomodulatory pathways following VARV infection.
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Use of live Variola Virus to determine whether CAST/EiJ mice are a suitable surrogate animal model for human smallpox
Virus research, 2019Co-Authors: Nadia Gallardo-romero, Victoria A. Olson, Scott K. Smith, Ashley V. Kondas, Darin S. Carroll, Christina L. Hutson, Johanna S. Salzer, Sharon Dietz-ostergaard, Paul Hudson, Inger K. DamonAbstract:Numerous animal models of systemic orthopoxVirus disease have been developed to evaluate therapeutics against Variola Virus (VARV), the causative agent of smallpox. These animal models do not resemble the disease presentation in human smallpox and most used surrogate OrthopoxViruses. A rodent model using VARV has a multitude of advantages, and previous investigations identified the CAST/EiJ mouse as highly susceptible to monkeypox Virus infection, making it of interest to determine if these rodents are also susceptible to VARV infection. In this study, we inoculated CAST/EiJ mice with a range of VARV doses (102-106 plaque forming units). Some animals had detectable viable VARV from the oropharynx between days 3 and 12 post inoculation. Despite evidence of disease, the CAST/EiJ mouse does not provide a model for clinical smallpox due to mild signs of morbidity and limited skin lesions. However, in contrast to previous rodent models using VARV challenge (i.e. prairie dogs and SCID mice), a robust immune response was observed in the CAST/EiJ mice (measured by Immunoglobulin G enzyme-linked immunosorbent assay). This is an advantage of this model for the study of VARV and presents a unique potential for the study of the immunomodulatory pathways following VARV infection.
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Variola Virus-Specific Diagnostic Assays: Characterization, Sensitivity, and Specificity
2016Co-Authors: Ashley V. Kondas, Victoria A. Olson, Richard Kline, Jason Abel, Miriam Laker, Laura Rose, Kimberly Wilkins, Jonathan Turner, Inger K. DamonAbstract:A public health response relies upon rapid and reliable confirmation of disease by diagnostic assays. Here, we detail the design and validation of two Variola Virus-specific real-time PCR assays, since previous assays cross-reacted with newly identified cow-pox Viruses. The assay specificity must continually be reassessed as other closely related Viruses are identified. Variola Virus, a solely human pathogen, is the causative agent ofsmallpox, which was deemed eradicated in 1980 by theWorld Health Organization (WHO), leading to the cessation of routine smallpox vaccination. Although several vaccines exist, the threat of Variola Virus being used as a bioweapon still remains (1, 2). It would be contraindicated for certain individuals, such as the im-munocompromised, to receive vaccination, due to serious vaccine adverse events that could be fatal (3, 4). Diagnosis based on clin-ical symptoms alone is challenging, since the symptoms of small-pox resemble those of numerous other rash-causing diseases and themajority of physicians today have never seen smallpox lesions. In the event of a release, quick and accurate clinical diagnosis would be critical to limit the spread of infection and contain
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Variola Virus-Specific Diagnostic Assays: Characterization, Sensitivity, and Specificity
Journal of clinical microbiology, 2015Co-Authors: Ashley V. Kondas, Victoria A. Olson, Richard Kline, Jason Abel, Miriam Laker, Kimberly Wilkins, Jonathan Turner, Laura J. Rose, Inger K. DamonAbstract:A public health response relies upon rapid and reliable confirmation of disease by diagnostic assays. Here, we detail the design and validation of two Variola Virus-specific real-time PCR assays, since previous assays cross-reacted with newly identified cowpox Viruses. The assay specificity must continually be reassessed as other closely related Viruses are identified.
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In Vitro Efficacy of Brincidofovir against Variola Virus
Antimicrobial Agents and Chemotherapy, 2014Co-Authors: Victoria A. Olson, E. Randall Lanier, Scott Foster, Scott K. Smith, Irina Gates, Lawrence C. Trost, Inger K. DamonAbstract:Brincidofovir (CMX001), a lipid conjugate of the acyclic nucleotide phosphonate cidofovir, is under development for smallpox treatment using “the Animal Rule,” established by the FDA in 2002. Brincidofovir reduces mortality caused by orthopoxVirus infection in animal models. Compared to cidofovir, brincidofovir has increased potency, is administered orally, and shows no evidence of nephrotoxicity. Here we report that the brincidofovir half-maximal effective concentration (EC50) against five Variola Virus strains in vitro averaged 0.11 μM and that brincidofovir was therefore nearly 100-fold more potent than cidofovir.
Scott K. Smith - One of the best experts on this subject based on the ideXlab platform.
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use of live Variola Virus to determine whether cast eij mice are a suitable surrogate animal model for human smallpox
Virus Research, 2020Co-Authors: Nadia F Gallardoromero, Victoria A. Olson, Scott K. Smith, Ashley V. Kondas, Darin S. Carroll, Christina L. Hutson, Johanna S. Salzer, Paul Hudson, Sharon Dietzostergaard, Inger K. DamonAbstract:Numerous animal models of systemic orthopoxVirus disease have been developed to evaluate therapeutics against Variola Virus (VARV), the causative agent of smallpox. These animal models do not resemble the disease presentation in human smallpox and most used surrogate OrthopoxViruses. A rodent model using VARV has a multitude of advantages, and previous investigations identified the CAST/EiJ mouse as highly susceptible to monkeypox Virus infection, making it of interest to determine if these rodents are also susceptible to VARV infection. In this study, we inoculated CAST/EiJ mice with a range of VARV doses (102-106 plaque forming units). Some animals had detectable viable VARV from the oropharynx between days 3 and 12 post inoculation. Despite evidence of disease, the CAST/EiJ mouse does not provide a model for clinical smallpox due to mild signs of morbidity and limited skin lesions. However, in contrast to previous rodent models using VARV challenge (i.e. prairie dogs and SCID mice), a robust immune response was observed in the CAST/EiJ mice (measured by Immunoglobulin G enzyme-linked immunosorbent assay). This is an advantage of this model for the study of VARV and presents a unique potential for the study of the immunomodulatory pathways following VARV infection.
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Use of live Variola Virus to determine whether CAST/EiJ mice are a suitable surrogate animal model for human smallpox
Virus research, 2019Co-Authors: Nadia Gallardo-romero, Victoria A. Olson, Scott K. Smith, Ashley V. Kondas, Darin S. Carroll, Christina L. Hutson, Johanna S. Salzer, Sharon Dietz-ostergaard, Paul Hudson, Inger K. DamonAbstract:Numerous animal models of systemic orthopoxVirus disease have been developed to evaluate therapeutics against Variola Virus (VARV), the causative agent of smallpox. These animal models do not resemble the disease presentation in human smallpox and most used surrogate OrthopoxViruses. A rodent model using VARV has a multitude of advantages, and previous investigations identified the CAST/EiJ mouse as highly susceptible to monkeypox Virus infection, making it of interest to determine if these rodents are also susceptible to VARV infection. In this study, we inoculated CAST/EiJ mice with a range of VARV doses (102-106 plaque forming units). Some animals had detectable viable VARV from the oropharynx between days 3 and 12 post inoculation. Despite evidence of disease, the CAST/EiJ mouse does not provide a model for clinical smallpox due to mild signs of morbidity and limited skin lesions. However, in contrast to previous rodent models using VARV challenge (i.e. prairie dogs and SCID mice), a robust immune response was observed in the CAST/EiJ mice (measured by Immunoglobulin G enzyme-linked immunosorbent assay). This is an advantage of this model for the study of VARV and presents a unique potential for the study of the immunomodulatory pathways following VARV infection.
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a multiplex pcr ldr assay for the simultaneous identification of category a infectious pathogens agents of viral hemorrhagic fever and Variola Virus
PLOS ONE, 2015Co-Authors: Sanchita Das, Victoria A. Olson, Scott K. Smith, Mark S. Rundell, Aashiq H. Mirza, Maneesh R. Pingle, Kristi Shigyo, Aura R. Garrison, Jason Paragas, Davise H. LaroneAbstract:CDC designated category A infectious agents pose a major risk to national security and require special action for public health preparedness. They include Viruses that cause viral hemorrhagic fever (VHF) syndrome as well as Variola Virus, the agent of smallpox. VHF is characterized by hemorrhage and fever with multi-organ failure leading to high morbidity and mortality. Smallpox, a prior scourge, has been eradicated for decades, making it a particularly serious threat if released nefariously in the essentially non-immune world population. Early detection of the causative agents, and the ability to distinguish them from other pathogens, is essential to contain outbreaks, implement proper control measures, and prevent morbidity and mortality. We have developed a multiplex detection assay that uses several species-specific PCR primers to generate amplicons from multiple pathogens; these are then targeted in a ligase detection reaction (LDR). The resultant fluorescently-labeled ligation products are detected on a universal array enabling simultaneous identification of the pathogens. The assay was evaluated on 32 different isolates associated with VHF (ebolaVirus, marburgVirus, Crimean Congo hemorrhagic fever Virus, Lassa fever Virus, Rift Valley fever Virus, Dengue Virus, and Yellow fever Virus) as well as Variola Virus and vaccinia Virus (the agent of smallpox and its vaccine strain, respectively). The assay was able to detect all Viruses tested, including 8 sequences representative of different Variola Virus strains from the CDC repository. It does not cross react with other emerging zoonoses such as monkeypox Virus or cowpox Virus, or six flaviViruses tested (St. Louis encephalitis Virus, Murray Valley encephalitis Virus, Powassan Virus, Tick-borne encephalitis Virus, West Nile Virus and Japanese encephalitis Virus).
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A Multiplex PCR/LDR Assay for the Simultaneous Identification of Category A Infectious Pathogens: Agents of Viral Hemorrhagic Fever and Variola Virus
2015Co-Authors: Sanchita Das, Victoria A. Olson, Scott K. Smith, Mark S. Rundell, Aashiq H. Mirza, Maneesh R. Pingle, Kristi Shigyo, Aura R. Garrison, Jason Paragas, Davise H. LaroneAbstract:CDC designated category A infectious agents pose a major risk to national security and require special action for public health preparedness. They include Viruses that cause viral hemorrhagic fever (VHF) syndrome as well as Variola Virus, the agent of smallpox. VHF is characterized by hemorrhage and fever with multi-organ failure leading to high morbidity and mortality. Smallpox, a prior scourge, has been eradicated for decades, making it a particularly serious threat if released nefariously in the essentially non-immune world population. Early detection of the causative agents, and the ability to distinguish them from other pathogens, is essential to contain outbreaks, implement proper control measures, and prevent morbidity and mortality. We have developed a multiplex detection assay that uses several species-specific PCR primers to generate amplicons from multiple pathogens; these are then targeted in a ligase detection reaction (LDR). The resultant fluorescently-labeled ligation products are detected on a universal array enabling simultaneous identification of the pathogens. The assay was evaluated on 32 different isolates associated with VHF (ebolaVirus, marburgVirus, Crimean Congo hemorrhagic fever Virus, Lassa fever Virus, Rift Valley fever Virus, Dengue Virus, and Yellow fever Virus) as well as Variola Virus and vaccinia Virus (the agent of smallpox and its vaccine strain, respectively). The assay was able to detect all Viruses tested, including 8 sequences representative of different Variola Virus strains from the CDC repository. It does not cross react with other emerging zoonoses such as monkeypox Virus or cowpox Virus, or six flaviViruses tested (St. Louis encephalitis Virus, Murray Valley encephalitis Virus, Powassan Virus, Tick-borne encephalitis Virus, West Nile Virus and Japanese encephalitis Virus).
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In Vitro Efficacy of Brincidofovir against Variola Virus
Antimicrobial Agents and Chemotherapy, 2014Co-Authors: Victoria A. Olson, E. Randall Lanier, Scott Foster, Scott K. Smith, Irina Gates, Lawrence C. Trost, Inger K. DamonAbstract:Brincidofovir (CMX001), a lipid conjugate of the acyclic nucleotide phosphonate cidofovir, is under development for smallpox treatment using “the Animal Rule,” established by the FDA in 2002. Brincidofovir reduces mortality caused by orthopoxVirus infection in animal models. Compared to cidofovir, brincidofovir has increased potency, is administered orally, and shows no evidence of nephrotoxicity. Here we report that the brincidofovir half-maximal effective concentration (EC50) against five Variola Virus strains in vitro averaged 0.11 μM and that brincidofovir was therefore nearly 100-fold more potent than cidofovir.
Victoria A. Olson - One of the best experts on this subject based on the ideXlab platform.
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use of live Variola Virus to determine whether cast eij mice are a suitable surrogate animal model for human smallpox
Virus Research, 2020Co-Authors: Nadia F Gallardoromero, Victoria A. Olson, Scott K. Smith, Ashley V. Kondas, Darin S. Carroll, Christina L. Hutson, Johanna S. Salzer, Paul Hudson, Sharon Dietzostergaard, Inger K. DamonAbstract:Numerous animal models of systemic orthopoxVirus disease have been developed to evaluate therapeutics against Variola Virus (VARV), the causative agent of smallpox. These animal models do not resemble the disease presentation in human smallpox and most used surrogate OrthopoxViruses. A rodent model using VARV has a multitude of advantages, and previous investigations identified the CAST/EiJ mouse as highly susceptible to monkeypox Virus infection, making it of interest to determine if these rodents are also susceptible to VARV infection. In this study, we inoculated CAST/EiJ mice with a range of VARV doses (102-106 plaque forming units). Some animals had detectable viable VARV from the oropharynx between days 3 and 12 post inoculation. Despite evidence of disease, the CAST/EiJ mouse does not provide a model for clinical smallpox due to mild signs of morbidity and limited skin lesions. However, in contrast to previous rodent models using VARV challenge (i.e. prairie dogs and SCID mice), a robust immune response was observed in the CAST/EiJ mice (measured by Immunoglobulin G enzyme-linked immunosorbent assay). This is an advantage of this model for the study of VARV and presents a unique potential for the study of the immunomodulatory pathways following VARV infection.
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Use of live Variola Virus to determine whether CAST/EiJ mice are a suitable surrogate animal model for human smallpox
Virus research, 2019Co-Authors: Nadia Gallardo-romero, Victoria A. Olson, Scott K. Smith, Ashley V. Kondas, Darin S. Carroll, Christina L. Hutson, Johanna S. Salzer, Sharon Dietz-ostergaard, Paul Hudson, Inger K. DamonAbstract:Numerous animal models of systemic orthopoxVirus disease have been developed to evaluate therapeutics against Variola Virus (VARV), the causative agent of smallpox. These animal models do not resemble the disease presentation in human smallpox and most used surrogate OrthopoxViruses. A rodent model using VARV has a multitude of advantages, and previous investigations identified the CAST/EiJ mouse as highly susceptible to monkeypox Virus infection, making it of interest to determine if these rodents are also susceptible to VARV infection. In this study, we inoculated CAST/EiJ mice with a range of VARV doses (102-106 plaque forming units). Some animals had detectable viable VARV from the oropharynx between days 3 and 12 post inoculation. Despite evidence of disease, the CAST/EiJ mouse does not provide a model for clinical smallpox due to mild signs of morbidity and limited skin lesions. However, in contrast to previous rodent models using VARV challenge (i.e. prairie dogs and SCID mice), a robust immune response was observed in the CAST/EiJ mice (measured by Immunoglobulin G enzyme-linked immunosorbent assay). This is an advantage of this model for the study of VARV and presents a unique potential for the study of the immunomodulatory pathways following VARV infection.
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Variola Virus-Specific Diagnostic Assays: Characterization, Sensitivity, and Specificity
2016Co-Authors: Ashley V. Kondas, Victoria A. Olson, Richard Kline, Jason Abel, Miriam Laker, Laura Rose, Kimberly Wilkins, Jonathan Turner, Inger K. DamonAbstract:A public health response relies upon rapid and reliable confirmation of disease by diagnostic assays. Here, we detail the design and validation of two Variola Virus-specific real-time PCR assays, since previous assays cross-reacted with newly identified cow-pox Viruses. The assay specificity must continually be reassessed as other closely related Viruses are identified. Variola Virus, a solely human pathogen, is the causative agent ofsmallpox, which was deemed eradicated in 1980 by theWorld Health Organization (WHO), leading to the cessation of routine smallpox vaccination. Although several vaccines exist, the threat of Variola Virus being used as a bioweapon still remains (1, 2). It would be contraindicated for certain individuals, such as the im-munocompromised, to receive vaccination, due to serious vaccine adverse events that could be fatal (3, 4). Diagnosis based on clin-ical symptoms alone is challenging, since the symptoms of small-pox resemble those of numerous other rash-causing diseases and themajority of physicians today have never seen smallpox lesions. In the event of a release, quick and accurate clinical diagnosis would be critical to limit the spread of infection and contain
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a multiplex pcr ldr assay for the simultaneous identification of category a infectious pathogens agents of viral hemorrhagic fever and Variola Virus
PLOS ONE, 2015Co-Authors: Sanchita Das, Victoria A. Olson, Scott K. Smith, Mark S. Rundell, Aashiq H. Mirza, Maneesh R. Pingle, Kristi Shigyo, Aura R. Garrison, Jason Paragas, Davise H. LaroneAbstract:CDC designated category A infectious agents pose a major risk to national security and require special action for public health preparedness. They include Viruses that cause viral hemorrhagic fever (VHF) syndrome as well as Variola Virus, the agent of smallpox. VHF is characterized by hemorrhage and fever with multi-organ failure leading to high morbidity and mortality. Smallpox, a prior scourge, has been eradicated for decades, making it a particularly serious threat if released nefariously in the essentially non-immune world population. Early detection of the causative agents, and the ability to distinguish them from other pathogens, is essential to contain outbreaks, implement proper control measures, and prevent morbidity and mortality. We have developed a multiplex detection assay that uses several species-specific PCR primers to generate amplicons from multiple pathogens; these are then targeted in a ligase detection reaction (LDR). The resultant fluorescently-labeled ligation products are detected on a universal array enabling simultaneous identification of the pathogens. The assay was evaluated on 32 different isolates associated with VHF (ebolaVirus, marburgVirus, Crimean Congo hemorrhagic fever Virus, Lassa fever Virus, Rift Valley fever Virus, Dengue Virus, and Yellow fever Virus) as well as Variola Virus and vaccinia Virus (the agent of smallpox and its vaccine strain, respectively). The assay was able to detect all Viruses tested, including 8 sequences representative of different Variola Virus strains from the CDC repository. It does not cross react with other emerging zoonoses such as monkeypox Virus or cowpox Virus, or six flaviViruses tested (St. Louis encephalitis Virus, Murray Valley encephalitis Virus, Powassan Virus, Tick-borne encephalitis Virus, West Nile Virus and Japanese encephalitis Virus).
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Variola Virus-Specific Diagnostic Assays: Characterization, Sensitivity, and Specificity
Journal of clinical microbiology, 2015Co-Authors: Ashley V. Kondas, Victoria A. Olson, Richard Kline, Jason Abel, Miriam Laker, Kimberly Wilkins, Jonathan Turner, Laura J. Rose, Inger K. DamonAbstract:A public health response relies upon rapid and reliable confirmation of disease by diagnostic assays. Here, we detail the design and validation of two Variola Virus-specific real-time PCR assays, since previous assays cross-reacted with newly identified cowpox Viruses. The assay specificity must continually be reassessed as other closely related Viruses are identified.
S. N. Shchelkunov - One of the best experts on this subject based on the ideXlab platform.
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The gene therapy of collagen-induced arthritis in rats by intramuscular administration of the plasmid encoding TNF-binding domain of Variola Virus CrmB protein.
Doklady. Biochemistry and biophysics, 2016Co-Authors: S. N. Shchelkunov, T. V. Tregubchak, O. S. Taranov, Rinat A. Maksyutov, Alexander N. Silkov, Sergey V. SennikovAbstract:Wistar rats with collagen-induced arthritis were intramuscularly injected with the recombinant plasmid pcDNA/sTNF-BD encoding the sequence of the TNF-binding protein domain of Variola Virus CrmB protein (VARV sTNF-BD) or the pcDNA3.1 vector. Quantitative analysis showed that the histopathological changes in the hind-limb joints of rats were most severe in the animals injected with pcDNA3.1 and much less severe in the group of rats injected with pcDNA/sTNF-BD, which indicates that gene therapy of rheumatoid arthritis is promising in the case of local administration of plasmids governing the synthesis of VARV immunomodulatory proteins.
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TNF binding protein of Variola Virus acts as a TNF antagonist at epicutaneous application.
Current pharmaceutical biotechnology, 2015Co-Authors: I. P. Gileva, S. N. Shchelkunov, Elena A. Viazovaia, Ludmila B. Toporkova, Dondok D. Tsyrendorzhiev, Irina A. OrlovskayaAbstract:VARV-CrmB is a TNF binding protein of Variola Virus. VARV-CrmB protein was previously shown to be active as a TNF-antagonist in a number of in vivo and in vitro models. Here we investigated the epicutaneous effect of recombinant VARV-CrmB protein using an experimental model of muTNFinduced migration of skin leukocytes as well as colony forming activity of bone marrow cells (BMC). Epiсutaneous applications of muTNF enhanced the number of cells migrating from skin flaps of BALB/c mice, whereas subsequent applications of VARV-CrmB protein in 30 min after muTNF, abolished that effect. Epicutaneously applied muTNF influenced the activity of committed hematopoietic progenitors causing a reduction of erythroid (BFUe+CFUe) colonies and increase of granulocyte-macrophage (CFU-GM) colonies in the colony-forming tests. VARV-CrmB, applied in combination with muTNF, demonstrated an ability to reverse this effect, namely, to increase BFUe+CFUe and reduce CFU-GM back to the control levels. Taking together, these data demonstrate the TNF-blocking properties of VARV-CrmB in vivo at epicutaneous applications. As effective TNF antagonist VARV-CrmB protein might be conceded as a beneficial candidate for future research and development of therapeutic approaches in the field of inflammatory skin diseases.
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SECRET domain of Variola Virus CrmB protein can be a member of poxviral type II chemokine-binding proteins family.
BMC research notes, 2010Co-Authors: Denis V. Antonets, Tatyana S Nepomnyashchikh, S. N. ShchelkunovAbstract:Variola Virus (VARV) the causative agent of smallpox, eradicated in 1980, have wide spectrum of immunomodulatory proteins to evade host immunity. Recently additional biological activity was discovered for VARV CrmB protein, known to bind and inhibit tumour necrosis factor (TNF) through its N-terminal domain homologous to cellular TNF receptors. Besides binding TNF, this protein was also shown to bind with high affinity several chemokines which recruit B- and T-lymphocytes and dendritic cells to sites of viral entry and replication. Ability to bind chemokines was shown to be associated with unique C-terminal domain of CrmB protein. This domain named SECRET (Smallpox Virus-Encoded Chemokine Receptor) is unrelated to the host proteins and lacks significant homology with other known viral chemokine-binding proteins or any other known protein. De novo modelling of VARV-CrmB SECRET domain spatial structure revealed its apparent structural homology with cowpox Virus CC-chemokine binding protein (vCCI) and vaccinia Virus A41 protein, despite low sequence identity between these three proteins. Potential ligand-binding surface of modelled VARV-CrmB SECRET domain was also predicted to bear prominent electronegative charge which is characteristic to known orthopoxviral chemokine-binding proteins. Our results suggest that SECRET should be included into the family of poxviral type II chemokine-binding proteins and that it might have been evolved from the vCCI-like predecessor protein.
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Recombinant TNF-binding protein from Variola Virus as a novel potential TNF antagonist.
Biochemistry. Biokhimiia, 2009Co-Authors: I. P. Gileva, T. S. Nepomnyashchikh, I. A. Ryazankin, S. N. ShchelkunovAbstract:Gel-filtration chromatographic separation of the lysate of Sf21 insect cells infected with recombinant baculoVirus BVi67 containing the gene for TNF-binding protein (CrmB) of Variola Virus (VARV) revealed that hTNF-cytotoxicity neutralization activity is associated with a fraction corresponding mainly to high molecular weight proteins (above 500 kDa) and less with fractions corresponding to proteins of 270 or 90 kDa. The recombinant VARV-CrmB protein has been purified by affinity chromatography. Difference in the experimentally determined and estimated (according to amino acid composition) VARV-CrmB molecular weight is due to glycosylation of the recombinant protein expressed in the insect cells. VARV-CrmB neutralizes in vitro the cytotoxic effect of hTNF and hLTα, and its TNF-neutralizing activity is two to three orders of magnitude higher compared to the analogous effects of type I and II soluble TNF receptors, comparable with the activity of mAb MAK195, and somewhat lower than the effect of the commercial drug Remicade.
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Comparative analysis of variable regions in the Variola Virus genome
Molecular Biology, 2008Co-Authors: I. V. Babkin, I. N. Babkina, T. S. Nepomnyashchikh, R. A. Maksyutov, S. N. ShchelkunovAbstract:Two segments of the variable terminal regions of the Variola Virus (VARV) genome were sequenced in 22 strains from the Russian collection, including about 13.5 kb of the left segment and about 10.5 kb of the right segment. The total length of the sequences was over 540 kb. Phylogenetic analysis of the new and published data determined the relationships among 70 VARV strains, the character of their clustering, and the intergroup and intragroup variation of the strain clusters. Loci with the highest polymorphism rate were identified and proved to map to noncoding regions or to regions of damaged open reading frames, characteristic of the ancestral Virus. These loci offer attractive possibilities for developing a strategy of VARV strain genotyping. Recombination analysis by different methods did not detect, except for a single case, significant recombination events in the VARV strains examined.
Roselyn J. Eisenberg - One of the best experts on this subject based on the ideXlab platform.
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characterization of chimpanzee human monoclonal antibodies to vaccinia Virus a33 glycoprotein and its Variola Virus homolog in vitro and in a vaccinia Virus mouse protection model
Journal of Virology, 2007Co-Authors: Zhaochun Chen, Scott K. Smith, Inger K. Damon, Roselyn J. Eisenberg, Patricia L. Earl, Jeffrey L. Americo, Andrew Sebrell, Suzanne U. Emerson, Gary H. Cohen, Inna GorshkovaAbstract:Three distinct chimpanzee Fabs against the A33 envelope glycoprotein of vaccinia Virus were isolated and converted into complete monoclonal antibodies (MAbs) with human γ1 heavy-chain constant regions. The three MAbs (6C, 12C, and 12F) displayed high binding affinities to A33 (Kd of 0.14 nM to 20 nM) and may recognize the same epitope, which was determined to be conformational and located within amino acid residues 99 to 185 at the C terminus of A33. One or more of the MAbs were shown to reduce the spread of vaccinia Virus as well as Variola Virus (the causative agent of smallpox) in vitro and to more effectively protect mice when administered before or 2 days after intranasal challenge with virulent vaccinia Virus than a previously isolated mouse anti-A33 MAb (1G10) or vaccinia Virus immunoglobulin. The protective efficacy afforded by anti-A33 MAb was comparable to that of a previously isolated chimpanzee/human anti-B5 MAb. The combination of anti-A33 MAb and anti-B5 MAb did not synergize the protective efficacy. These chimpanzee/human anti-A33 MAbs may be useful in the prevention and treatment of vaccinia Virus-induced complications of vaccination against smallpox and may also be effective in the immunoprophylaxis and immunotherapy of smallpox and other orthopoxVirus diseases.
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Characterization of chimpanzee/human monoclonal antibodies to vaccinia Virus A33 glycoprotein and its Variola Virus homolog in vitro and in a vaccinia Virus mouse protection model.
Journal of virology, 2007Co-Authors: Zhaochun Chen, Scott K. Smith, Inger K. Damon, Patricia L. Earl, Jeffrey L. Americo, Andrew Sebrell, Suzanne U. Emerson, Gary H. Cohen, Roselyn J. EisenbergAbstract:Three distinct chimpanzee Fabs against the A33 envelope glycoprotein of vaccinia Virus were isolated and converted into complete monoclonal antibodies (MAbs) with human γ1 heavy-chain constant regions. The three MAbs (6C, 12C, and 12F) displayed high binding affinities to A33 (Kd of 0.14 nM to 20 nM) and may recognize the same epitope, which was determined to be conformational and located within amino acid residues 99 to 185 at the C terminus of A33. One or more of the MAbs were shown to reduce the spread of vaccinia Virus as well as Variola Virus (the causative agent of smallpox) in vitro and to more effectively protect mice when administered before or 2 days after intranasal challenge with virulent vaccinia Virus than a previously isolated mouse anti-A33 MAb (1G10) or vaccinia Virus immunoglobulin. The protective efficacy afforded by anti-A33 MAb was comparable to that of a previously isolated chimpanzee/human anti-B5 MAb. The combination of anti-A33 MAb and anti-B5 MAb did not synergize the protective efficacy. These chimpanzee/human anti-A33 MAbs may be useful in the prevention and treatment of vaccinia Virus-induced complications of vaccination against smallpox and may also be effective in the immunoprophylaxis and immunotherapy of smallpox and other orthopoxVirus diseases.
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Major neutralizing sites on vaccinia Virus glycoprotein B5 are exposed differently on Variola Virus ortholog B6.
Journal of virology, 2007Co-Authors: Lydia Aldaz-carroll, Yuhong Xiao, J. Charles Whitbeck, Manuel Ponce De Leon, Huan Lou, Mikyung Kim, Ellis L. Reinherz, Stuart N. Isaacs, Roselyn J. EisenbergAbstract:Immunization against smallpox (Variola Virus) with Dryvax, a live vaccinia Virus (VV), was effective, but now safety is a major concern. To overcome this issue, subunit vaccines composed of VV envelope proteins from both forms of infectious virions, including the extracellular enveloped virion (EV) protein B5, are being developed. However, since B5 has 23 amino acid differences compared with its B6 Variola Virus homologue, B6 might be a better choice for such a strategy. Therefore, we compared the properties of both proteins using a panel of monoclonal antibodies (MAbs) to B5 that we had previously characterized and grouped according to structural and functional properties. The B6 gene was obtained from the Centers for Disease Control and Prevention, and the ectodomain was cloned and expressed in baculoVirus as previously done with B5, allowing us to compare the antigenic properties of the proteins. Polyclonal antibodies to B5 or B6 cross-reacted with the heterologous protein, and 16 of 26 anti-B5 MAbs cross-reacted with B6. Importantly, 10 anti-B5 MAbs did not cross-react with B6. Of these, three have important anti-VV biologic properties, including their ability to neutralize EV infectivity and block comet formation. Here, we found that one of these three MAbs protected mice from a lethal VV challenge by passive immunization. Thus, epitopes that are present on B5 but not on B6 would generate an antibody response that would not recognize B6. Assuming that B6 contains similar Variola Virus-specific epitopes, our data suggest that a subunit vaccine using the Variola Virus homologues might exhibit improved protective efficacy against smallpox.
