A naturally occurring cowpox virus with an ectromelia virus A-type inclusion protein gene displays atypical A-type inclusions

Okeke, Malachy Ifeanyi; Hansen, Hilde; Traavik, Terje

doi:10.1016/j.meegid.2011.09.017

Cited by 21 publications

(42 citation statements)

References 49 publications

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“…Among OPVs, only CPXV, ECTV and RCNV are known to produce ATI in infected cells [22,32,33]. Three strain specific ATI phenotypes are known to exist; the V + ATI in which virions are within the ATI matrix, the V − phenotype in which the ATI lacks virions within or on the surface, and the V +/ phenotype in which the ATIs lack internalized virions but are encrusted with virions on their surface [24,32,33]. To determine the ATI phenotype produced in cells infected with Fennoscandian CPXVs, we quantified ATI phenotypes in 50 cell sections that were clearly infected.…”

Section: Resultsmentioning

confidence: 99%

“…The same ATI phenotype was produced in A549 cells (data not shown). Electron micrographs of the V + ATI produced in CPXV-No-H1 infected cells and the V +/ ATI produced in CPXV-No-H2 infected cells have been published previously [24,33]. Bars: 5 μm (A-F) .…”

Section: Resultsmentioning

confidence: 99%

“…The XbaI restriction enzyme digestion of PCR products amplified with ATI-2 primer pairs [30,33] has proved to be robust in identifying and differentiating OPVs [30]. To differentiate Fennoscandian CPXVs, we amplified a fragment of the atip gene with ATI-2 primer pairs (Figure 2A) and digested the obtained PCR products for 2 hours with XbaI (Figure 2B).…”

Section: Resultsmentioning

confidence: 99%

“…The CPXV-No-H2 restriction pattern is similar to that of ECTV-MOS. Characterization of CPXV-No-H2 as a naturally occurring recombinant between ECTV and CPXV has been reported elsewhere [33]. …”

Section: Resultsmentioning

confidence: 99%

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Molecular characterization and phylogenetics of Fennoscandian cowpox virus isolates based on the p4c and atip genes

Okeke

Okoli

Nilssen

et al. 2014

Virol J

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BackgroundCowpox virus (CPXV), a rodent-borne Orthopoxvirus (OPV) that is indigenous to Eurasia can infect humans, cattle, felidae and other animals. Molecular characterization of CPXVs isolated from different geographic locations is important for the understanding of their biology, geographic distribution, classification and evolution. Our aim was to characterize CPXVs isolated from Fennoscandia on the basis of A-type inclusion (ATI) phenotype, restriction fragment length polymorphism (RFLP) profiles of atip gene fragment amplicon, and phylogenetic tree topology in conjunction with the patristic and genetic distances based on full length DNA sequence of the atip and p4c genes.MethodsATI phenotypes were determined by transmission electron microcopy and RFLP profiles were obtained by restriction enzyme digestion of the atip gene fragment PCR product. A 6.2 kbp region spanning the entire atip and p4c genes of Fennoscandian CPXV isolates was amplified and sequenced. The phylogenetic affinity of Fennoscandian CPXV isolates to OPVs isolated from other geographic regions was determined on the basis of the atip and p4c genes.ResultsFennoscandian CPXV isolates encoded full length atip and p4c genes. They produce wild type V+ ATI except for CPXV-No-H2. CPXVs were resolved into six and seven species clusters based on the phylogeny of the atip and p4c genes respectively. The CPXVs isolated from Fennoscandia were grouped into three distinct clusters that corresponded to isolates from Norway, Sweden and Finland.ConclusionCPXV is a polyphyletic assemblage of six or seven distinct clusters and the current classification in which CPXVs are united as one single species should be re-considered. Our results are of significance to the classification and evolution of OPVs.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Molecular characterization and phylogenetics of Fennoscandian cowpox virus isolates based on the p4c and atip genes

Okeke

Okoli

Nilssen

et al. 2014

Virol J

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“…Most natural CPXV strains produce V ϩ or V Ϫ ATIs as the dominant but stable phenotype. Recently, the first poxvirus isolate, CPXV-No-H2, was described that produced V ϩ/ ATI bodies as a stable phenotype (48). Nevertheless, all other CPXVs isolated from Fennoscandia represent the V ϩ ATI phenotype and, hence, encode full-length ati and p4c genes (49).…”

Section: Discussionmentioning

confidence: 99%

Out of the Reservoir: Phenotypic and Genotypic Characterization of a Novel Cowpox Virus Isolated from a Common Vole

Hoffmann

Franke

Jenckel

et al. 2015

J Virol

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The incidence of human cowpox virus (CPXV) infections has increased significantly in recent years. Serological surveys have suggested wild rodents as the main CPXV reservoir. We characterized a CPXV isolated during a large-scale screening from a feral common vole. A comparison of the full-length DNA sequence of this CPXV strain with a highly virulent pet rat CPXV isolate showed a sequence identity of 96%, including a large additional open reading frame (ORF) of about 6,000 nucleotides which is absent in the reference CPXV strain Brighton Red. Electron microscopy analysis demonstrated that the vole isolate, in contrast to the rat strain, forms A-type inclusion (ATI) bodies with incorporated virions, consistent with the presence of complete ati and p4c genes. Experimental infections showed that the vole CPXV strain caused only mild clinical symptoms in its natural host, while all rats developed severe respiratory symptoms followed by a systemic rash. In contrast, common voles infected with a high dose of the rat CPXV showed severe signs of respiratory disease but no skin lesions, whereas infection with a low dose led to virus excretion with only mild clinical signs. We concluded that the common vole is susceptible to infection with different CPXV strains. The spectrum ranges from well-adapted viruses causing limited clinical symptoms to highly virulent strains causing severe respiratory symptoms. In addition, the low pathogenicity of the vole isolate in its eponymous host suggests a role of common voles as a major CPXV reservoir, and future research will focus on the correlation between viral genotype and phenotype/ pathotype in accidental and reservoir species. IMPORTANCEWe report on the first detection and isolation of CPXV from a putative reservoir host, which enables comparative analyses to understand the infection cycle of these zoonotic orthopox viruses and the relevant genes involved. In vitro studies, including whole-genome sequencing as well as in vivo experiments using the Wistar rat model and the vole reservoir host allowed us to establish links between genomic sequences and the in vivo properties (virulence) of the novel vole isolate in comparison to those of a recent zoonotic CPXV isolated from pet rats in 2009. Furthermore, the role of genes present only in a reservoir isolate can now be further analyzed. These studies therefore allow unique insights and conclusions about the role of the rodent reservoir in CPXV epidemiology and transmission and about the zoonotic threat that these viruses represent. C owpox virus (CPXV), a member of the genus Orthopoxvirus (OPV) in the Poxviridae family, is suspected to be widespread in Western Eurasian rodents, particularly vole species (1, 2). From the presumed reservoir hosts, spill-over infections to accidental hosts are regularly observed (3). The accidental hosts include domestic cats and also exotic animals in zoos, such as large felids and elephants, which regularly develop severe disease (3). As CPXV is a zoonotic virus, humans in direct contact wit...

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Molecular investigation and cultivation of camelpox virus in Iran

et al. 2014

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Camelpox virus (genus Orthopoxvirus, family Poxviridae) is the etiologic agent of camel pox. The clinical manifestations of this virus range from inapparent infection to mild, moderate and, less commonly, severe systemic infection and death. Following an outbreak of camelpox, samples that were collected from camel flocks suspected to have camelpox in Qom Province in central Iran and Khash city, Sistan and Baluchestan Province and South Khorasan Province in eastern Iran were sent to Razi Vaccine and Serum Research Institute in Mashhad. DNA extraction was performed primarily by the phenol-chloroform method, and PCR was carried out using a Bioneer kit. Using the primer pair 5'-AAT-ACA-AGG-AGG-ATC-T-3' and 5'-CTT-AAC-TTT-TTC-TTT-CTC-3', the gene sequence encoding the A-type inclusion protein (ATIP) was amplified. The size of the PCR product, specific for camelpox virus, was 881 bp. The PCR product was purified, and to confirm its sequence, it was sent to the reference laboratory. The sequence was subjected to a BLAST search and then phylogenetically analyzed using CLC software. The results showed that all samples were nearly 100 % identical to each other and to strains CMS and M-96. These isolates also had 99 % and 95 % similarity to the CP-1 strain and isolate FIN/T2000, respectively. In Vero cell culture, inoculation with this virus caused a cytopathic effect (CPE), which appeared 2-5 days post-inoculation. Characteristic CPE showing foci of rounded cells, ballooning, giant-cell formation and syncytia with degenerative changes appeared.

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A naturally occurring cowpox virus with an ectromelia virus A-type inclusion protein gene displays atypical A-type inclusions

Cited by 21 publications

References 49 publications

Molecular characterization and phylogenetics of Fennoscandian cowpox virus isolates based on the p4c and atip genes

Molecular characterization and phylogenetics of Fennoscandian cowpox virus isolates based on the p4c and atip genes

Out of the Reservoir: Phenotypic and Genotypic Characterization of a Novel Cowpox Virus Isolated from a Common Vole

Molecular investigation and cultivation of camelpox virus in Iran

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