Virus Interferon Resistance Research Articles

Detection and phylogenetic analysis of Orf virus in Kashmir Himalayas.

Orf virus (ORFV) is a zoonotic pathogen that primarily infects sheep and goats, and is responsible for significant economic losses. ORFV is endemic in all the major sheep and goat rearing areas of the world including Indian subcontinent. However, the nature of ORFV circulating among sheep and goat in Kashmir Himalayas has not yet been characterized. In the present study, we describe natural outbreaks of ORFV in sheep and goats of Kashmir Himalayas. We detected the presence of ORFV in the scab lesion by PCR amplification of the major envelope protein (B2L) gene. We sequenced the virus interferon resistance (VIR) gene and determined their phylogenetic relationship with that of the published reference sequences. Phylogenetic analysis based on VIR gene revealed that the ORFV isolates from Kashmir Himalayas separated into main two clusters. The sheep isolates showed genetic homology with the sheep strains reported from Greece and Italy, whereas the goat-specific strain show homology with the goat strains reported from China. This study demonstrates the presence of ORFV infection in sheep and goats, and report first phylogenetic analysis of the ORFV strains prevalent in the Kashmir Himalayas.

Molecular characterization of Orf virus in goats in Gabon, Central Africa.

BackgroundOrf or contagious ecthyma is a zoonotic viral infection with a potential serious health threat for the small ruminants industry as well as humans. It is currently emerging in new territories.ResultsEight suspected clinical cases of pustular dermatitis in goats occurred in the rural area of Tebe, in south-eastern Gabon, in January 2013. The orf virus (ORFV) was detected by high-throughput sequencing on sera, buccal swabs and scab pool samples. It was confirmed in six out of eight sick goats by using specific PCR targeting the major envelope protein (B2L) and the orf virus interferon resistance (VIR) genes. Phylogenetic analysis revealed that the Gabonese strain and South Korean strains evolved from a common ancestor, suggesting an Asian origin of the ORFV’ Gabonese strain.ConclusionsThis study provides the molecular detection of the ORFV strain involved in the cases of pustular dermatitis in goats and highlights its circulation in Gabon.

Phylogenetic analysis of two Chinese orf virus isolates based on sequences of B2L and VIR genes

Orf virus (ORFV) is an enveloped virus with a double-stranded DNA genome, causing a contagious pustular dermatitis, mainly in goats and sheep. In this study, two strains of ORFV were isolated from sheep and goat samples in Xinjiang and Shaanxi, China. The B2L and virus interferon resistance (VIR) genes of these two isolates were sequenced and analyzed after PCR amplification. Phylogenetic analysis showed that the two isolates clustered with other ORFV strains but were separated into different subgroups. The Xinjiang strain shared the highest homology with the Gansu strain, whereas the Shaanxi strain shared higher homology with the Taiwan and Hubei strains. This is the first report of the molecular characterization of ORFV in Northwest China, and it provides new information on the genotyping of the causative agents responsible for contagious ecthyma dermatitis outbreaks in China.

Isolation and characterization of orf viruses from Korean black goats

Five cases of orf virus infection in Korean black goats were diagnosed in our laboratory between 2010 and 2011. One orf virus (ORF/2011) was isolated from an ovine testis cell line (OA3.Ts) for use as a vaccine candidate. Sequences of the major envelope protein and orf virus interferon resistance genes were determined and compared with published reference sequences. Phylogenetic analyses revealed that orf viruses from Korean black goats were most closely related to an isolate (ORF/09/Korea) from dairy goats in Korea. This result indicates that the orf viruses might have been introduced from dairy goats into the Korean black goat population.

Looking back: a genetic retrospective study of Brazilian Orf virus isolates

Orf virus (ORFV), the prototype of the genus Parapoxvirus, is the aetiological agent of contagious ecthyma (CE), a pustular dermatitis that afflicts domestic and wild small ruminants. CE is one...

Spatial and Temporal Genetic Homogeneity of Orf Viruses Infecting Japanese Serows (Capricornis crispus)

This study genetically characterized orf viruses (ORFVs) isolated from recent outbreaks in Japanese serows (Capricornis crispus) in 2007 and 2008, and from earlier outbreaks from 1985 to 2001. Nucleotide sequences of genes for the viral envelope, vascular endothelial growth factor (VEGF), and virus interferon resistance (VIR) were determined in two ORFVs isolated from recent outbreaks and in eight from earlier outbreaks. No deletions or insertions were observed in these genes. Surprisingly, the amino acid sequences of the envelope and VIR genes were identical among the 10 ORFVs, and only one amino acid substitution in the VEGF gene was found in one of the two recent ORFVs (2007). Genotyping by differential PCR for the A32L gene, which encodes an ATPase, classified all of the 10 ORFVs into the same genotype. In an ORFV isolated from a sheep in 1970, the three sequenced genes were almost the same as in the ORFVs isolated from Japanese serows, and the ORFV in 1970 was classified into the same genotype as the ORFVs from Japanese serows. These results suggest that recent outbreaks in Japanese serows are caused by ORFVs genetically closely related to the viruses in earlier outbreaks and that these genetically stable ORFVs have circulated among Japanese serows over a long period of time.

Phylogenetic analysis and characterization of Korean orf virus from dairy goats: case report

An outbreak of orf virus infection in dairy goats in Korea was investigated. Suspected samples of the skin and lip of affected goats were sent to the laboratory for more exact diagnosis. Orf virus was detected by electron microscopy and viral DNA was identified by PCR. To reveal the genetic characteristics of the Korean strain (ORF/09/Korea), the sequences of the major envelope protein (B2L) and orf virus interferon resistance (VIR) genes were determined and then compared with published reference sequences. Phylogenetic analysis revealed that the ORF/09/Korea strain was closest to the isolates (Taiping) from Taiwan. This is believed to be the first report on the molecular characterization of orf virus in Korea.

The N-terminal domain of the vaccinia virus E3L-protein is required for neurovirulence, but not induction of a protective immune response

Encephalitis is a rare, but serious complication from vaccination against smallpox using replication competent strains of vaccinia virus. In this report we describe mutants of vaccinia virus, containing N-terminal deletions of the vaccinia virus interferon resistance gene, E3L, that are attenuated for neuropathogenesis in a mouse model system. These recombinant viruses replicated to high titers in the nasal mucosa after intra-nasal infection of C57BL/6 mice but failed to spread to the lungs or brain. These viruses demonstrated reduced pathogenicity after intra-cranial infection as well, indicating a decrease in neurovirulence. Intra-nasal inoculation or inoculation by scarification with a low dose of recombinant virus containing a deletion of the entire N-terminal domain of E3L protected against challenge with a high dose of wild-type vaccinia virus, suggesting that this replication competent, but attenuated strain of vaccinia virus may have promise as an improved vaccine for protecting against smallpox, and as a vector for inducing mucosal immunity to heterologous pathogenic organisms.

Orf virus immuno-modulation and the host immune response

Orf virus encodes a range of immuno-modulatory genes that interfere with host anti-virus immune and inflammatory effector mechanisms. The function of these reflects the pathogenesis of orf. The orf virus interferon resistance protein (OVIFNR) and virus IL-10 (vIL-10) inhibit interferon production and activity. In addition the vIL-10 suppresses inflammatory cytokine production by activated macrophages and keratinocytes. The virus GM-CSF inhibitory factor (GIF) is a novel virus protein that binds to and inhibits the biological activity of GM-CSF and IL-2. Together, these immuno-modulators target key effector mechanisms of host anti-virus immunity to allow time for virus replication in epidermal cells.

Both carboxy- and amino-terminal domains of the vaccinia virus interferon resistance gene, E3L, are required for pathogenesis in a mouse model.

The vaccinia virus (VV) E3L gene is responsible for providing interferon (IFN) resistance and a broad host range to VV in cell culture. The E3L gene product contains two distinct domains. A conserved carboxy-terminal domain, which is required for the IFN resistance and broad host range of the virus, has been shown to bind double-stranded RNA (dsRNA) and inhibit the antiviral dsRNA-dependent protein kinase, PKR. The amino-terminal domain, while conserved among orthopoxviruses, is dispensable in cell culture. To study the role of E3L in whole-animal infections, WR strain VV recombinants either lacking E3L (VVDeltaE3L) or expressing an amino-terminal (VVE3LDelta83N) or carboxy-terminal (VVE3LDelta26C) truncation of E3L were constructed. Whereas wild-type VV had a 50% lethal dose of approximately 10(4) PFU after intranasal infection, and elicited severe weight loss and morbidity, VVDeltaE3L was apathogenic, leading to no death, weight loss, or morbidity. VVDeltaE3L was also apathogenic after intracranial injection. Although the amino-terminal domain of E3L is dispensable for infection of cells in culture, both the amino- and carboxy-terminal domains of E3L were required for full pathogenesis in intranasal infections. These results demonstrate that the entire E3L gene is required for pathogenesis in the mouse model.

Orf virus encodes a homolog of the vaccinia virus interferon-resistance gene E3L.

A homolog of the vaccinia virus (VAC) interferon resistance gene E3L has been discovered in orf virus strain NZ-2, a parapoxvirus that infects sheep, goats and humans. The gene is located 20 kb from the left terminus of the orf virus genome and is transcribed towards this terminus. RNase protection studies have been used to define the limits of the gene and Northern analysis revealed that it is expressed early in infection. The predicted amino acid sequence of the orf virus protein shares 31% identity (57% similarity) with the VAC E3L protein. Four of the six residues identified as being essential to dsRNA binding in the vaccinia virus protein are conserved in the orf virus protein whilst the other two amino acid changes are conservative substitutions. The orf virus gene has been sequenced in two other orf virus strains which vary markedly in their ability to produce experimental lesions in vivo. Their predicted protein sequences vary by less than 3% from the NZ-2 protein. The recombinant orf virus protein, expressed as a fusion protein in E. coli, bound double-stranded (ds)RNA but not dsDNA, single-stranded (ss)DNA or ssRNA . This is the first demonstration of a VAC E3L-like gene encoded by a parapoxvirus.