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At least 19 recordsLinked to original sources

Poxvirus orthologous clusters: toward defining the minimum essential poxvirus genome.

Increasingly complex bioinformatic analysis is necessitated by the plethora of sequence information currently available. A total of 21 poxvirus genomes have now been completely sequenced and annotated, and many more genomes will be available in the next few years. First, we describe the creation of a database of continuously corrected and updated genome sequences and an easy-to-use and extremely powerful suite of software tools for the analysis of genomes, genes, and proteins. These tools are available free to all researchers and, in most cases, alleviate the need for using multiple Internet sites for analysis. Further, we describe the use of these programs to identify conserved families of genes (poxvirus orthologous clusters) and have named the software suite POCs, which is available at www.poxvirus.org. Using POCs, we have identified a set of 49 absolutely conserved gene families-those which are conserved between the highly diverged families of insect-infecting entomopoxviruses and vertebrate-infecting chordopoxviruses. An additional set of 41 gene families conserved in chordopoxviruses was also identified. Thus, 90 genes are completely conserved in chordopoxviruses and comprise the minimum essential genome, and these will make excellent drug, antibody, vaccine, and detection targets. Finally, we describe the use of these tools to identify necessary annotation and sequencing updates in poxvirus genomes. For example, using POCs, we identified 19 genes that were widely conserved in poxviruses but missing from the vaccinia virus strain Tian Tan 1998 GenBank file. We have reannotated and resequenced fragments of this genome and verified that these genes are conserved in Tian Tan. The results for poxvirus genes and genomes are discussed in light of evolutionary processes.

Amino Acid Sequence↗

Reticuloendotheliosis virus (REV) long terminal repeats incorporated in the genomes of commercial fowl poxvirus vaccines and pigeon poxviruses without indication of the presence of infectious REV.

Because of reticuloendotheliosis virus (REV) contamination in commercial poultry vaccines, polymerase chain reaction (PCR) assays have been described to increase the sensitivity of biological assays used to detect REV in vaccines. The PCR assay designed to amplify the long terminal repeat (LTR) region of REV identified REV LTRs in many of the commercial fowl poxvirus (FPV) vaccines evaluated. These commercial vaccines were not thought to be contaminated with replicating REV because of the lack of REV outbreaks, the lack of in vitro amplification, and lack of a serologic response to REV. As previously described, the FPV S vaccine strain is known to carry infectious integrated proviral REV, whereas FPV M vaccine strain and its derivatives carry integrated LTRs or remnants of REV proviral DNA inserted into the FPV genome. Another PCR assay designed to amplify the envelope gene of REV was used to verify that the envelope proviral gene was not present in REV LTR PCR-positive samples. Southern blot analysis with REV LTR probes hybridized to the 9-kb EcoRI genomic fragment of all FPV and pigeon poxviruses evaluated, whereas the envelope probe did not hybridize to any poxvirus genome. Sequence analysis of the 9-kb EcoRI fragment indicated that an integrated REV LTR exists in the 9-kb EcoRI of some poxvirus genomes. A new PCR assay designed to amplify integrated REV LTRs in the 9-kb EcoRI fragment identified complete and incomplete integrated REV LTRs in all FPV and pigeon poxvirus genomes evaluated.

Animals↗

Poxvirus in scaled quail and prevalences of poxvirus-like lesions in northern bobwhites and scaled quail from Texas.

Prevalences of poxvirus-like lesions were determined for 177 northern bobwhites (Colinus virginianus) and 24 scaled quail (Callipepla squamata) trapped in southern Texas from 1976 to 1979 and for 190 northern bobwhites and 105 scaled quail shot at five locations in southern Texas from 1980 to 1981. None of the northern bobwhites trapped in 1976-1977 was infected, but 54% of the trapped scaled quail were infected; 17% of the northern bobwhites and 34% of the scaled quail shot in 1980-1981 had pox lesions, primarily on the wings. Prevalence was unrelated to sex or age of birds. For both species, prevalence was greatest during late spring and early summer. Histologic and electron microscopic examination confirmed poxvirus in two scaled quail, which constituted the first report of poxvirus in this species.

Age Factors↗

The white pock (mu) mutants of rabbit poxvirus. III. Terminal DNA sequence duplication and transposition in rabbit poxvirus.

The restriction fragment patterns of the DNA of three white pock (mu) nonhost range mutants RPmu mutants) of rabbit poxvirus (RPV) show the presence of net insertions of DNA. Two of the mutants (RPmu21 and RPmu7) contain two molar quantities of 13 X 10(6) and 15 X 10(6) daltons, respectively, of the left-most viral DNA sequences. The extra copy of these sequences was inserted into the right-most region of the viral genome which in both cases had been modified by deletion of 10 X 10(6) daltons of the original extreme right-most DNA sequences. Hence two mutants with "left-hand" mirror image termini of 13 X 10(6) and 15 X 10(6) daltons of DNA were generated with an overall net increase in mass of 3 X 10(6) and 5 X 10(6) daltons of DNA to yield genome molecular weights of 121 X 10(6) and 123 X 10(6) for RPmu21 and 7, respectively. A third mutant (RPmu81g) contained 28 X 10(6) daltons of the right-most parental DNA sequences inserted into the left end of the genome, which had deleted 6 X 10(6) daltons of the extreme left-hand sequences. This variant contains "right-hand" mirror image termini of 28 X 10(6) daltons of DNA and a net increase in molecular weight of 22 X 10(6) daltons of DNA to yield a genome molecular weight of 140 X 10(6). The cross-linked end of the transposed sequences is retained in all cases. Our data suggest that the terminal DNA sequences at either end of the virus are not essential for a productive infection in certain host cells and that the pathogenicity of orthopoxviruses may be related in part to a series of varied and interchangeable DNA sequences located at both extremes of a high conserved genome core.

Base Sequence↗

Interferon induction in peripheral blood mononuclear leukocytes of man and farm animals by poxvirus vector candidates and some poxvirus constructs.

Prototypes of three poxvirus genera--orthopoxvirus (OPV), parapoxvirus (PPV), avipoxvirus (APV)--and Newcastle disease virus (NDV) as a control, as well as three recombinant OPV strains and one recombinant APV strain, were incubated in vitro with peripheral blood mononuclear leukocytes (PBML) of man, sheep and swine. Antiviral activity was determined in PBML culture supernatants at different time intervals after virus cell interaction using a cytopathic effect inhibition bioassay. Additionally, supernatants derived from human PBML were screened for interferons (IFN) alpha and gamma as well as for tumor necrosis factor by enzyme-linked immunosorbent assay. IFN titers reached a maximum 24 h after PBML stimulation at a multiplicity of infection (MOI) greater than 1. IFN alpha/beta was found to be responsible for the antiviral effect. Using a MOI > or = 1 the highly attenuated strain MVA was the only representant of vaccinia virus (VV) that induced significant amounts of IFN also as a lacZ recombinant. Replicable virus from five well-known VV strains as well as the Chinese VV strain Tien Tan (VVTT) as a recombinant vaccine failed to induce leukocyte IFN. Inactivated VV strain Elstree and the recombinant TT strain induced high titers of leukocyte IFN. Supernatants derived from human, porcine and ovine PBML stimulated with replicable PPV, native VV MVA and MVA lacZ recombinant or native APV and APV lacZ recombinant virus regularly contained IFN alpha. In contrast to NDV, neither specific antisera nor monoclonal antibodies were able to block the INF induction by VV and PPV.

Animals↗

An Evolutionary Framework Exploiting Virologs and Their Host Origins to Inform Poxvirus Protein Functions.

Poxviruses represent evolutionary successful infectious agents. As a family, poxviruses can infect a wide variety of species including humans, fish, and insects. While many other viruses are species-specific, an individual poxvirus species is often capable of infecting diverse hosts and cell types. For example, the prototypical poxvirus, vaccinia, is well known to infect numerous human cell types but can also infect cells from divergent hosts like frog neurons. Notably, poxvirus infections result in both detrimental human and animal diseases. The most infamous disease linked to a poxvirus is smallpox caused by variola virus. Poxviruses are large double-stranded DNA viruses, which uniquely replicate in the cytoplasm of cells. The model poxvirus genome encodes ~200 nonoverlapping protein-coding open reading frames (ORFs). Poxvirus gene products impact various biological processes like the production of virus particles, the host range of infectivity, and disease pathogenesis. In addition, poxviruses and their gene products have biomedical application with several species commonly engineered for use as vaccines and oncolytic virotherapy. Nevertheless, we still have an incomplete understanding of the functions associated with many poxvirus genes. In this chapter, we outline evolutionary insights that can complement ongoing studies of poxvirus gene functions and biology, which may serve to elucidate new molecular activities linked to this biomedically relevant class of viruses.

Animals↗

Immune response to poxvirus infections in various animals.

The study of infections of vertebrate animals by poxviruses has remained a dynamic area of research for the last century. The host range of poxviruses vary from extremely narrow to exceedingly broad, and they have been shown to enter their host by either the respiratory route or through the skin. The severity of infection varies dramatically from one species to another, causing anywhere from a local, self-limiting infection, to a devastating systemic disease, such as smallpox. Although the immune response to poxvirus infections are very similar to that seen in other viral infections, the poxviruses, unlike most other viruses (with the exception of Herpes viruses), are able to defend themselves. They have been shown to carry a repertoire of proteins involved in immune evasion and immune modulation. Poxviruses encode proteins involved in blocking many of the strategies employed by the host to combat viral infections; they encode for proteins that block activity of many chemokines, cytokines, serine proteases, and even complement. Traditionally, different animal models have been used to study the pathogenesis of poxvirus infections, and the characterization of virulence genes using mutant poxviruses. Additionally, new animal models are being developed to study the possible therapeutic uses many of these poxvirus immune modulating proteins might have. This review discusses the host immune response against poxvirus infections in various animals, the viral counter response to the host, and the animal models used to study poxvirus infection and immune modulating proteins.

Animals↗

Genetic identification of novel poxviruses of cetaceans and pinnipeds.

Novel poxviruses were identified in skin lesions of several species of cetaceans and pinnipeds using polymerase chain reaction targeting DNA polymerase and DNA topoisomerase I genes of members of the subfamily Chordopoxvirinae. With the exception of parapoxviruses, no molecular data of marine mammal poxviruses were available to infer genetic and evolutionary relatedness to terrestrial vertebrate poxviruses. Viruses were assigned to a cetacean poxvirus 1 (CPV-1) group based on nucleotide and amino acid identities of gene fragments amplified from skin lesions of Asian bottlenose (Tursiops aduncus), Atlantic bottlenose (Tursiops truncatus), rough-toothed (Steno bredanensis), and striped (Stenella coeruleoalba) dolphins. A different poxvirus was detected in skin lesions of a bowhead whale (Balaena mysticetus) and provisionally assigned to a CPV-2 group. These viruses showed highest identity to terrestrial poxviruses of the genera Orthopoxvirus and Suipoxvirus. A novel species-specific poxvirus was also identified in skin lesions of Steller sea lions (Eumetopias jubatus). None of these poxviruses were found to have amplifiable hemagglutinin gene sequences. Novel parapoxviruses were also identified in skin lesions of Steller sea lions and spotted seals (Phoca largha). A significant degree of divergence was observed in sequences of Steller sea lion parapoxviruses, while those of spotted seals and harbor seals (Phoca vitulina) were highly conserved.

Alaska↗

Zoonotic poxvirus infections in humans.

PURPOSE OF REVIEW: The 2003 USA monkeypox epidemic caused by imported African rodents, newly emergent poxvirus zoonoses in Brazil and the possible use of variola virus for biological warfare has led to renewed interest in poxviruses and anti-poxviral therapies. Increasing foreign travel and importation of exotic animal species increases the likelihood of poxvirus infections occurring outside their usual geographical range and diagnostic delay has important implications. The present review provides an overview of these rare zoonoses. RECENT FINDINGS: Three genera of Poxviridae are known to cause human zoonoses: orthopoxviruses, parapoxviruses and yatapoxvirus. Most cases are occupational, sporadic and have few cutaneous lesions with low morbidity. The exception is monkeypox, similar to smallpox, with significant morbidity and childhood mortality. Molecular characterization using polymerase chain reaction (PCR) amplification and other methods provides accurate phylogenetic identification and suggests that a cowpox-like virus is the probable ancestor of variola and other zoonotic poxviruses. DNA genomic sequencing of the Brazilian Cantagalo and Araçatuba viruses shows a close relationship to vaccinia virus. Poxviruses have potential in cancer immunotherapy and their ability to evade host-cell immune responses may provide a basis for new antipoxvirus therapies. Other agents, particularly nucleoside phosphonates such as cidofovir, show therapeutic action against poxviruses. SUMMARY: Human zoonotic poxvirus infections are rare but increasingly encountered outside their usual geographical range. The 2003 USA monkeypox outbreak emphasizes the importance of early accurate diagnosis, particularly because increasing numbers of immunosuppressed individuals increases the potential for severe or fatal infections. PCR methodology enables accurate phylogenetic typing and has identified new diseases, but rapid, reliable methods must be made available for clinicians. More research into therapeutic agents for the prevention and treatment of poxvirus infections is required.

Animals↗

Genetic and antigenic characterization of a poxvirus isolate from ostriches.

Avian poxvirus was isolated from nodules on the heads and conjunctiva of two 3-to-4-wk-old ostrich chicks. The ostriches from which poxvirus was isolated had been placed on premises where turkeys that had shown evidence of poxvirus infection had been raised earlier. Microscopically, the nodules from the ostriches were composed of proliferating and hypertrophic epithelial cells that formed large fronds. Most of the hypertrophic epithelial cells contained large eosinophilic intracytoplasmic inclusion bodies characteristic of poxvirus. Characterization of the avian poxvirus isolated from the cutaneous lesions in ostriches was based on western blotting of virus antigen, restriction fragment length polymorphism of genomic DNA, pathogenesis, and cross-protection studies in chickens. Antigenic and genetic studies did not reveal any significant difference between the poxvirus isolated from ostriches (PVO) and fowl poxvirus (FPV). Further, susceptible chickens immunized with the PVO were protected when challenged with a virulent strain of FPV. Thus, the poxvirus isolated from ostriches had similar antigenic, genetic, and biological properties to FPV.

Animals↗

F-box-like domains are present in most poxvirus ankyrin repeat proteins.

Vertebrate poxviruses encode numerous proteins with the ankyrin (ANK) repeat, protein-protein interaction motif but little is known about the role(s) of this large family of poxvirus proteins. We report here that the vast majority of poxvirus ANK repeat proteins share a general molecular architecture that includes a conserved amino acid motif at the carboxyl terminus. This motif is most like the F-box seen in a range of cellular proteins. From 80-100% of the ANK repeat proteins of any one poxvirus have an F-box-like domain and we observed only one poxvirus protein with an F-box-like domain but lacking ANK repeats. The proteins of only one genus of vertebrate poxviruses lack F-box-like domains and this genus does not encode ANK repeat proteins. Many F-box proteins are recognition subunits of ubiquitin ligase complexes in which the F-box binds to core elements of the complex and protein-protein interaction domains in the remainder of the protein bind the substrate protein. These observations suggest a general model of the function of the poxvirus ANK-F-box proteins. We propose that the F-box-like domains in these proteins interact with cellular ubiquitin ligase complexes and thereby direct the ubiquitination of proteins bound to the ANK repeats. The large number of different poxviral ANK-F-box proteins suggests a wide range of cellular proteins might be subjected to ubiquitin-mediated degradation, thereby modulating diverse cellular responses to viral infection.

Amino Acid Sequence↗

Poxviruses: past, present and future.

The analysis of poxvirus genomes is complex, in part, because of their size (130-360 kb) and the fact that gene content is variable; a common set of 49 genes has been found in all sequenced poxviruses and an additional 41 genes are also present in all sequenced orthopoxviruses. As a group, poxviruses have a very broad range of eukaryotic hosts (including mammals, birds, reptiles and insects) and many poxvirus genes are associated with blocking host anti-viral responses. One consequence of this is that many poxvirus genes are not essential for growth in tissue culture and that extensive passaging in vitro results in the accumulation of mutations, including deletions that result in loss of gene function. Here, we review various comparative analyses of the poxviruses including gene prediction, gene conservation and function, genome organization, and poxvirus taxonomy and evolution.

Animals↗

Poxvirus tropism.

Despite the success of the WHO-led smallpox eradication programme a quarter of a century ago, there remains considerable fear that variola virus, or other related pathogenic poxviruses such as monkeypox, could re-emerge and spread disease in the human population. Even today, we are still mostly ignorant about why most poxvirus infections of vertebrate hosts show strict species specificity, or how zoonotic poxvirus infections occur when poxviruses occasionally leap into novel host species. Poxvirus tropism at the cellular level seems to be regulated by intracellular events downstream of virus binding and entry, rather than at the level of specific host receptors as is the case for many other viruses. This review summarizes our current understanding of poxvirus tropism and host range, and discusses the prospects of exploiting host-restricted poxvirus vectors for vaccines, gene therapy or tissue-targeted oncolytic viral therapies for the treatment of human cancers.

Animals↗