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PLoS Pathogens Oct 2018The recent de novo assembly of horsepox is an instructive example of an information hazard: published methods enabling poxvirus synthesis led to media coverage spelling...
The recent de novo assembly of horsepox is an instructive example of an information hazard: published methods enabling poxvirus synthesis led to media coverage spelling out the implications, efficiently disseminating true information that might be used to cause harm. Whether or not the benefits justified the risks, the horsepox saga provides ample reason to upgrade the current system for screening synthesized DNA for hazardous sequences, which does not cover the majority of firms and cannot reliably prevent the assembly of potentially pandemic pathogens. An upgraded system might leverage one-way encryption to confidentially scrutinize virtually all commercial production by a cooperative international network of servers whose integrity can be verified by third parties. Funders could support participating institutions to ease the transition or outright subsidize the market to make clean DNA cheaper, while boycotts by journals, institutions, and funders could ensure compliance and require hardware-level locks on future DNA synthesizers. However, the underlying problem is that security and safety discussions among experts typically follow potentially hazardous events rather than anticipating them. Changing norms and incentives to favor preregistration and advisory peer review of planned experiments could test alternatives to the current closeted research model in select areas of science. Because the fields of synthetic mammalian virology and especially gene drive research involve technologies that could be unilaterally deployed and may self-replicate in the wild, they are compelling candidates for initial trials of early-stage peer review.
Topics: Biohazard Release; Biomedical Research; Humans; Information Dissemination; Orthopoxvirus; Pandemics; Poxviridae Infections; Risk Assessment; Safety Management
PubMed: 30286188
DOI: 10.1371/journal.ppat.1007286 -
Journal of Virology Oct 2017The castaneous (CAST) mouse, a wild-derived inbred strain, is highly susceptible to orthopoxvirus infection by intranasal and systemic routes. The 50% lethal...
The castaneous (CAST) mouse, a wild-derived inbred strain, is highly susceptible to orthopoxvirus infection by intranasal and systemic routes. The 50% lethal intraperitoneal dose of vaccinia virus (VACV) was 3 PFU for CAST mice, whereas BALB/c mice survived 10 PFU. At all times and in all organs analyzed, virus titers were higher in CAST than in BALB/c mice. In individual CAST mice, luciferase-expressing VACV was seen to replicate rapidly leading to death, whereas virus levels increased for a few days and then declined in BALB/c mice. Increases in gamma interferon (IFN-γ) and tumor necrosis factor alpha (TNF-α) were delayed and low in CAST mice compared to BALB/c mice following VACV infection or poly(I-C) inoculation, consistent with differences in innate immune responses. In addition, naive CAST mice had considerably lower numbers of NK and T cells than BALB/c mice. The percentage of IFN-γ-producing CD4 and CD8 T cells increased following infection of CAST mice only after considerable virus spread, and the absolute cell numbers remained low. Administration of exogenous IFN-γ or -α to CAST mice before or during the first days of infection suppressed virus replication and prolonged survival, allowing the mice to make adaptive CD4 and CD8 T cell responses that were necessary to clear the virus after cessation of interferon treatment. Thus, insufficient innate cytokine and cellular immune responses contribute to the unique susceptibility of CAST mice to VACV, whereas the adaptive immune response can be protective only if virus replication is suppressed during the first several days of infection. Most inbred mouse strains are relatively resistant to orthopoxviruses. The castaneous (CAST) mouse is a notable exception, exhibiting extreme vulnerability to monkeypox virus, cowpox virus, and vaccinia virus and thus providing a unique model for studying pathogenicity, immunity, vaccines, and antiviral drugs. To fully utilize the CAST mouse for such purposes, it is necessary to understand the basis for virus susceptibility. We showed that naive CAST mice make low IFN-γ and TNF-α responses and have low levels of NK cells and CD4 and CD8 T cells compared to a resistant classical inbred mouse strain. Attenuating virus replication with one or more doses of exogenous IFN-α or -γ before or during the first few days of infection enabled the development of adaptive cellular immunity and clearance of virus. Further genetic studies may reveal the basis for the low innate immunity.
Topics: Animals; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Line; Chlorocebus aethiops; Female; Immunity, Innate; Interferon-gamma; Killer Cells, Natural; Lymphocyte Count; Mice; Mice, Inbred BALB C; Poxviridae Infections; Tumor Necrosis Factor-alpha; Vaccinia virus; Virus Replication
PubMed: 28747505
DOI: 10.1128/JVI.01042-17 -
Microbiology and Immunology 2002The possibility of persistent parapoxvirus (PPV) infection was investigated by serologically and genetically using cattle infected with the virus experimentally and...
The possibility of persistent parapoxvirus (PPV) infection was investigated by serologically and genetically using cattle infected with the virus experimentally and naturally. Three cattle were inoculated with the virus subcutaneously at several spots in the lips and abdominal regions. Small papules developed in the inoculated regions, and antibodies to the virus developed and continued persistently. One animal, from which one PPV had been previously isolated, was also subjected to serological and viral detection tests as a naturally infected case. Two of these four cattle were injected with dexamethasone (DM), and one was injected with interferon-gamma (IFN-gamma). The viral genome was rarely detected from the peripheral blood leukocytes in the ordinary condition, but frequently when the animals were injected with IFN-gamma. The viral genome was also detected from the lymph nodes as these PPV infected animals were euthanized. These results indicated that cattle were infected with PPV subclinically and persistently, and the virus was activated in stressed or immunosuppressed animals. The virus would be harbored in the lymphotic tissues of the animals when they show no clinical symptoms.
Topics: Animals; Antibodies, Viral; Blotting, Southern; Cattle; Cattle Diseases; Cells, Cultured; DNA, Viral; Enzyme-Linked Immunosorbent Assay; Immunocompetence; Interferon-gamma; Parapoxvirus; Polymerase Chain Reaction; Poxviridae Infections
PubMed: 12061631
DOI: 10.1111/j.1348-0421.2002.tb02697.x -
Diseases of Aquatic Organisms Oct 1999Viruses belonging to 9 families have been detected in cetaceans. We critically review the clinical features, pathology and epidemiology of the diseases they cause.... (Review)
Review
Viruses belonging to 9 families have been detected in cetaceans. We critically review the clinical features, pathology and epidemiology of the diseases they cause. Cetacean morbillivirus (family Paramyxoviridae) induces a serious disease with a high mortality rate and persists in several populations. It may have long-term effects on the dynamics of cetacean populations either as enzootic infection or recurrent epizootics. The latter presumably have the more profound impact due to removal of sexually mature individuals. Members of the family Poxviridae infect several species of odontocetes, resulting in ring and tattoo skin lesions. Although poxviruses apparently do not induce a high mortality, circumstancial evidence suggests they may be lethal in young animals lacking protective immunity, and thus may negatively affect net recruitment. Papillomaviruses (family Papovaviridae) cause genital warts in at least 3 species of cetaceans. In 10% of male Burmeister's porpoises Phocoena spinipinnis from Peru, lesions were sufficiently severe to at least hamper, if not impede, copulation. Members of the families Herpesviridae, Orthomyxoviridae and Rhabdoviridae were demonstrated in cetaceans suffering serious illnesses, but with the exception of a 'porpoise herpesvirus' their causative role is still tentative. Herpes-like viruses and caliciviruses (Caliciviridae) give rise to cutaneous diseases in Monodontidae and Delphinidae. Antibodies to several serotypes of caliciviruses were found in odontocetes and mysticetes. An unrecognized Hepadnaviridae was detected by serology in a captive Pacific white-sided dolphin Lagenorhynchus obliquidens with chronic persistent hepatitis. Adenoviruses (Adenoviridae) were isolated from the intestinal tracts of mysticeti and a beluga Delphinapterus leucas but were not associated with any pathologies. We discuss the potential impact of Paramyxoviridae, Poxviridae and Papovaviridae on the dynamics of several odontocete populations.
Topics: Adenoviridae; Animals; Caliciviridae; Cetacea; Hepadnaviridae; Herpesviridae; Male; Morbillivirus; Morbillivirus Infections; Orthomyxoviridae; Papillomaviridae; Papillomavirus Infections; Poxviridae; Poxviridae Infections; Rhabdoviridae; Tumor Virus Infections
PubMed: 10590929
DOI: 10.3354/dao038053 -
Emerging Infectious Diseases Sep 2018We report detection and full-genome characterization of a novel orthopoxvirus (OPXV) responsible for a fatal infection in a cat. The virus induced skin lesions...
We report detection and full-genome characterization of a novel orthopoxvirus (OPXV) responsible for a fatal infection in a cat. The virus induced skin lesions histologically characterized by leukocyte infiltration and eosinophilic cytoplasmic inclusions. Different PCR approaches were unable to assign the virus to a defined OPXV species. Large amounts of typical brick-shaped virions, morphologically related to OPXV, were observed by electron microscopy. This OPXV strain (Italy_09/17) was isolated on cell cultures and embryonated eggs. Phylogenetic analysis of 9 concatenated genes showed that this virus was distantly related to cowpox virus, more closely related to to ectromelia virus, and belonged to the same cluster of an OPXV recently isolated from captive macaques in Italy. Extensive epidemiologic surveillance in cats and rodents will assess whether cats are incidental hosts and rodents are the main reservoir of the virus. The zoonotic potential of this novel virus also deserves further investigation.
Topics: Animals; Cat Diseases; Cats; Diagnosis, Differential; Fatal Outcome; Italy; Male; Orthopoxvirus; Poxviridae Infections
PubMed: 30124195
DOI: 10.3201/eid2409.171283 -
Veterinary Medicine and Science Aug 2019Peste-des-petits-ruminants (PPR) and Goat pox (GTP) are two devastating and economically important transboundary animal diseases of small ruminants in Africa and Asia...
Peste-des-petits-ruminants (PPR) and Goat pox (GTP) are two devastating and economically important transboundary animal diseases of small ruminants in Africa and Asia that have been difficult to control. This study however, investigated an outbreak of PPR and GTP in a mixed flock of indigenous sheep and goats in Kanam, North Central Nigeria. A total of nine sera and seven tissues (lungs, spleen, scab and skin) samples were collected and analysed in the laboratory using competitive enzyme linked immunosorbent assay (cELISA) for PPR antibodies and polymerase chain reaction (PCR) for detection of PPR virus (PPRV) and GTP virus (GTPV). Gene fragments of the nucleoprotein of PPRV and the G-protein-coupled chemokine receptor (GPCR) of GTPV were amplified and sequenced to confirm the presence of the causative viruses. Serologically, antibodies to PPRV were detected in all (9/9) sera collected. GTPV and PPRV was detected in corresponding samples (42.8% n = 3/7) of the scab/skin samples collected by both PCR and RT-PCR technique. The phylogenetic analysis of PPRV revealed that the virus belongs to lineage IV and clustered with viruses from Gabon and Cameroon. Similarly, the GTPV also clustered with other sequences from Burkina Faso and Yemen. The positive cELISA, RT-PCR and PCR results from samples collected from the same animals confirmed co-infection of PPR and GTP in this mixed flock of sheep and goats. This is the first report of concurrent infection of PPR and GTP in mixed flock of sheep and goats in Nigeria. Our findings underscore the need for farmers to vaccinate their flock to control spread and economic losses as result of these diseases.
Topics: Animals; Capripoxvirus; Coinfection; Disease Outbreaks; Goat Diseases; Goats; Nigeria; Peste-des-Petits-Ruminants; Peste-des-petits-ruminants virus; Phylogeny; Poxviridae Infections; Sheep; Sheep Diseases
PubMed: 30993915
DOI: 10.1002/vms3.170 -
Biochemical and Biophysical Research... Nov 2014Small nuclear RNAs (snRNAs) are <200 nucleotide non-coding uridylate-rich RNAs. Although the functions of many snRNAs remain undetermined, a population of snRNAs is...
Small nuclear RNAs (snRNAs) are <200 nucleotide non-coding uridylate-rich RNAs. Although the functions of many snRNAs remain undetermined, a population of snRNAs is produced during the early phase of infection of cells by vaccinia virus. In the present study, we demonstrate a direct correlation between expression of the cytoprotective enzyme heme oxygenase-1 (HO-1), suppression of selective snRNA expression, and inhibition of vaccinia virus infection of macrophages. Hemin induced HO-1 expression, completely reversed virus-induced host snRNA expression, and suppressed vaccinia virus infection. This involvement of specific virus-induced snRNAs and associated gene clusters suggests a novel HO-1-dependent host-defense pathway in poxvirus infection.
Topics: Gene Expression; Heme Oxygenase-1; Host-Pathogen Interactions; Humans; Immunity, Innate; Macrophages; Multigene Family; Poxviridae Infections; RNA, Small Nuclear; RNA, Untranslated; Vaccinia virus
PubMed: 25450361
DOI: 10.1016/j.bbrc.2014.10.035 -
Annals of the New York Academy of... Feb 2019The role of interferon (IFN)-induced protein kinase R (PKR) in capripoxvirus (CaPV)-infected cells remains unknown. In this study, we show that CaPV infection triggered...
The role of interferon (IFN)-induced protein kinase R (PKR) in capripoxvirus (CaPV)-infected cells remains unknown. In this study, we show that CaPV infection triggered PKR and eukaryotic translation initiation factor 2 alpha (eIF2α) protein phosphorylation in a dose-dependent manner, and that this leads to decreased CaPV replication. Overexpression of PKR compromised viral gene expression and inhibited sheeppox virus (SPPV) replication. Downregulation of PKR with siRNAs significantly decreased eIF2α phosphorylation and reduced the mRNA level of IFN-β, which increased virus replication. In luciferase assays, species-different CaPVs K3L proteins inhibited sheep PKR (sPKR): goatpox virus K3L strongly inhibited sPKR and goat PKR (gPKR), but SPPV K3L only partially inhibited gPKR. These results are the first to show that SPPV infection induces phosphorylation of eIF2α through PKR activation, which then results in restriction of CaPV replication. Furthermore, our data show that CaPV K3L inhibits PKR in a species-specific manner. The results presented are consistent with the hypothesis that different levels of PKR inhibition by K3L orthologs from various viruses could potentially contribute to the host range function of K3L.
Topics: Animals; Capripoxvirus; Cell Line, Tumor; Chlorocebus aethiops; Eukaryotic Initiation Factor-2; Goats; HeLa Cells; Humans; Interferon-gamma; Phosphorylation; Poxviridae Infections; RNA Interference; RNA, Small Interfering; Sheep; Vero Cells; Viral Proteins; Virus Replication; eIF-2 Kinase
PubMed: 30381842
DOI: 10.1111/nyas.13976 -
Biophysical Journal Oct 1997Single living cells were studied in growth medium by atomic force microscopy at a high--down to one image frame per second--imaging rate over time periods of many hours,... (Review)
Review
Single living cells were studied in growth medium by atomic force microscopy at a high--down to one image frame per second--imaging rate over time periods of many hours, stably producing hundreds of consecutive scans with a lateral resolution of approximately 30-40 nm. The cell was held by a micropipette mounted onto the scanner-piezo as shown in Häberle, W., J. K. H. Hörber, and G. Binnig. 1991. Force microscopy on living cells. J. Vac. Sci. Technol. B9:1210-0000. To initiate specific processes on the cell surface the cells had been infected with pox viruses as reported earlier and, most likely, the liberation of a progeny virion by the still-living cell was observed, hence confirming and supporting earlier results (Häberle, W., J. K. H. Hörber, F. Ohnesorge, D. P. E. Smith, and G. Binnig. 1992. In situ investigations of single living cells infected by viruses. Ultramicroscopy. 42-44:1161-0000; Hörber, J. K. H., W. Häberle, F. Ohnesorge, G. Binnig, H. G. Liebich, C. P. Czerny, H. Mahnel, and A. Mayr. 1992. Investigation of living cells in the nanometer regime with the atomic force microscope. Scanning Microscopy. 6:919-930). Furthermore, the pox viruses used were characterized separately by AFM in an aqueous environment down to the molecular level. Quasi-ordered structural details were resolved on a scale of a few nm where, however, image distortions and artifacts due to multiple tip effects are probably involved--just as in very high resolution (<15-20 nm) images on the cells. Although in a very preliminary manner, initial studies on the mechanical resonance properties of a single living (noninfected) cell, held by the micropipette, have been performed. In particular, frequency response spectra were recorded that indicate elastic properties and enough stiffness of these cells to make the demonstrated rapid scanning of the imaging tip plausible. Measurements of this kind, especially if they can be proven to be cell-type specific, may perhaps have a large potential for biomedical applications. Images of these living cells were also recorded in the widely known (e.g., Radmacher, M., R. W. Tillmann, and H. E. Gaub. 1993. Imaging viscoelasticity by force modulation with the atomic force microscope. Biophys. J. 64:735-742) force modulation mode, yet at one low modulation frequency of approximately 2 kHz. (Note: After the cells were attached to the pipette by suction, they first deformed significantly and then reassumed their original spherical shape, which they also acquire when freely suspended in solution, to a great extent with the exception of the portion adjusting to the pipette edge geometry after approximately 0.5-1 h, which occurred in almost the same manner with uninfected cells, and those that had been infected several hours earlier. This seems to be a process which is at least actively supported by the cellular cytoskeleton, rather than a mere osmotic pressure effect induced by electrolyte transport through the membrane. Furthermore, several hours postinfection (p.i.) infected cells developed many optically visible refraction effects, which appeared as small dark spots in the light microscope, that we believed to be the regions in the cell plasma where viruses are assembled; this is known from the literature on electron microscopy on pox-infected cells and referred to there as "virus factories" (e.g., Moss, B. 1986. Replication of pox viruses. In Fundamental Virology, B. N. Fields and D. M. Knape, editors. Raven Press, New York. 637-655). Therefore, we assume that the cells stay alive during imaging, in our experience for approximately 30-45 h p.i.).
Topics: Animals; Biomechanical Phenomena; Biophysical Phenomena; Biophysics; Cells, Cultured; Elasticity; Haplorhini; Microscopy, Atomic Force; Poxviridae; Poxviridae Infections; Virus Replication; Viscosity
PubMed: 9336215
DOI: 10.1016/S0006-3495(97)78250-X -
Vaccine Mar 2012The modified vaccinia virus Ankara (MVA) strain is a highly attenuated strain of vaccinia virus that has been demonstrated to be safe for humans. MVA is widely... (Review)
Review
The modified vaccinia virus Ankara (MVA) strain is a highly attenuated strain of vaccinia virus that has been demonstrated to be safe for humans. MVA is widely considered as the vaccinia virus strain of choice for clinical investigation because of its high safety profile. It also represents an excellent candidate for use as vector system in recombinant vaccine development for gene delivery or vaccination against infectious diseases or tumours, even in immunocompromised individuals. The use of MVA and recombinant MVA vectors must comply with various regulatory requirements, particularly relating to the assessment of potential risks for human health and the environment. The purpose of the present paper is to highlight some biological characteristics of MVA and MVA-based recombinant vectors and to discuss these from a biosafety point of view in the context of the European regulatory framework for genetically modified organisms with emphasis on the assessment of potential risks associated with environmental release.
Topics: Animals; Clinical Trials as Topic; Gene Transfer Techniques; Genetic Therapy; Genetic Vectors; Humans; Mammals; Neoplasms; Poxviridae Infections; Practice Guidelines as Topic; Vaccination; Vaccinia virus; Viral Vaccines
PubMed: 22342706
DOI: 10.1016/j.vaccine.2012.02.016