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Suppression of influenza virus infection by the orf virus isolated in Taiwan.

Lin FY, Tseng YY, Chan KW, Kuo ST, Yang CH, Wang CY, Takasu M, Hsu WL, Wong ML - J. Vet. Med. Sci. (2015)

Bottom Line: ORFV stimulated human monocytes (THP-1) secreting proinflammatory cytokines IL-8 and TNF-α.Similarly, mice infected with ORFV via both intramuscular and subcutaneous routes at two days prior to IAV infection significantly decreased the replication of IAV.In summary, the results of a current study indicated our Hoping strain harbors the immune modulator property; with such a bio-adjuvanticity, we further proved that pre-exposure of ORFV protects animals from subsequent IAV infection.

View Article: PubMed Central - PubMed

Affiliation: Department of Veterinary Medicine, College of Veterinary Medicine, National Chung Hsing University, Taichung 402, Taiwan.

ABSTRACT
Orf virus (ORFV), a member of parapoxvirus, is an enveloped virus with genome of double-stranded DNA. ORFV causes contagious pustular dermatitis or contagious ecthyma in sheep and goats worldwide. In general, detection of viral DNA and observing ORFV virion in tissues of afflicted animals are two methods commonly used for diagnosis of orf infection; however, isolation of the ORFV in cell culture using virus-containing tissue as inoculum is known to be difficult. In this work, the ORFV (Hoping strain) isolated in central Taiwan was successfully grown in cell culture. We further examined the biochemical characteristic of our isolate, including viral genotyping, viral mRNA and protein expression. By electron microscopy, one unique form of viral particle from ORFV infected cellular lysate was demonstrated in the negative-stained field. Moreover, immunomodulating and anti-influenza virus properties of this ORFV were investigated. ORFV stimulated human monocytes (THP-1) secreting proinflammatory cytokines IL-8 and TNF-α. And, pre-treatment of ORFV-infected cell medium prevents A549 cells from subsequent type A influenza virus (IAV) infection. Similarly, mice infected with ORFV via both intramuscular and subcutaneous routes at two days prior to IAV infection significantly decreased the replication of IAV. In summary, the results of a current study indicated our Hoping strain harbors the immune modulator property; with such a bio-adjuvanticity, we further proved that pre-exposure of ORFV protects animals from subsequent IAV infection.

No MeSH data available.


Related in: MedlinePlus

Cytopathic effect on primary goat testis cells caused by cellular lysate containingORFVs. (A) The infected primary goat testis cells showed rounding, shrinking anddetachment, and eventually formed a viral plaque (200 × magnification). Mock is theimage of uninfected cells. (B) Detection of orf viral DNA by PCR. The isolatedplaque (v) showed the expected sizes (~900 bp, as indicated by the arrow) ofamplified products of partial B2L gene. The positive control (+) wasthe virus-infected cell lysate in PCR; the negative control (–) wasdone without any DNA template. M is DNA size markers. (C) Identification of isolatedORFVs by the single-step PCR. As the PCR amplification yielded two DNA fragmentswith characteristic sizes (180 and 254 bp, as indicated by arrows), it indicated theisolated ORFV is the Hoping strain (Chan et al., 2009). (D)Patterns of orf viral DNA after restriction enzymes digestion. Lane 1: DNA treatedwith EcoR I; lane 2: treatment with BamH I; lane 3: treatment with Hind III; lane 4:treatment with Kpn I; lane 5 is the uninfected cell, and the stained DNA is smearingafter Kpn I digestion. M is the DNA size markers.
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fig_001: Cytopathic effect on primary goat testis cells caused by cellular lysate containingORFVs. (A) The infected primary goat testis cells showed rounding, shrinking anddetachment, and eventually formed a viral plaque (200 × magnification). Mock is theimage of uninfected cells. (B) Detection of orf viral DNA by PCR. The isolatedplaque (v) showed the expected sizes (~900 bp, as indicated by the arrow) ofamplified products of partial B2L gene. The positive control (+) wasthe virus-infected cell lysate in PCR; the negative control (–) wasdone without any DNA template. M is DNA size markers. (C) Identification of isolatedORFVs by the single-step PCR. As the PCR amplification yielded two DNA fragmentswith characteristic sizes (180 and 254 bp, as indicated by arrows), it indicated theisolated ORFV is the Hoping strain (Chan et al., 2009). (D)Patterns of orf viral DNA after restriction enzymes digestion. Lane 1: DNA treatedwith EcoR I; lane 2: treatment with BamH I; lane 3: treatment with Hind III; lane 4:treatment with Kpn I; lane 5 is the uninfected cell, and the stained DNA is smearingafter Kpn I digestion. M is the DNA size markers.

Mentions: Plaque purification of the ORFV: With continued viral passages, theinfected primary goat testis cells began to show the cytopathic effect (CPE) and form aviral plaque. The CPE in primary goat testis cells was local and limited on the area ofaffected cells after four to five days after infection (Fig. 1AFig. 1.


Suppression of influenza virus infection by the orf virus isolated in Taiwan.

Lin FY, Tseng YY, Chan KW, Kuo ST, Yang CH, Wang CY, Takasu M, Hsu WL, Wong ML - J. Vet. Med. Sci. (2015)

Cytopathic effect on primary goat testis cells caused by cellular lysate containingORFVs. (A) The infected primary goat testis cells showed rounding, shrinking anddetachment, and eventually formed a viral plaque (200 × magnification). Mock is theimage of uninfected cells. (B) Detection of orf viral DNA by PCR. The isolatedplaque (v) showed the expected sizes (~900 bp, as indicated by the arrow) ofamplified products of partial B2L gene. The positive control (+) wasthe virus-infected cell lysate in PCR; the negative control (–) wasdone without any DNA template. M is DNA size markers. (C) Identification of isolatedORFVs by the single-step PCR. As the PCR amplification yielded two DNA fragmentswith characteristic sizes (180 and 254 bp, as indicated by arrows), it indicated theisolated ORFV is the Hoping strain (Chan et al., 2009). (D)Patterns of orf viral DNA after restriction enzymes digestion. Lane 1: DNA treatedwith EcoR I; lane 2: treatment with BamH I; lane 3: treatment with Hind III; lane 4:treatment with Kpn I; lane 5 is the uninfected cell, and the stained DNA is smearingafter Kpn I digestion. M is the DNA size markers.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591145&req=5

fig_001: Cytopathic effect on primary goat testis cells caused by cellular lysate containingORFVs. (A) The infected primary goat testis cells showed rounding, shrinking anddetachment, and eventually formed a viral plaque (200 × magnification). Mock is theimage of uninfected cells. (B) Detection of orf viral DNA by PCR. The isolatedplaque (v) showed the expected sizes (~900 bp, as indicated by the arrow) ofamplified products of partial B2L gene. The positive control (+) wasthe virus-infected cell lysate in PCR; the negative control (–) wasdone without any DNA template. M is DNA size markers. (C) Identification of isolatedORFVs by the single-step PCR. As the PCR amplification yielded two DNA fragmentswith characteristic sizes (180 and 254 bp, as indicated by arrows), it indicated theisolated ORFV is the Hoping strain (Chan et al., 2009). (D)Patterns of orf viral DNA after restriction enzymes digestion. Lane 1: DNA treatedwith EcoR I; lane 2: treatment with BamH I; lane 3: treatment with Hind III; lane 4:treatment with Kpn I; lane 5 is the uninfected cell, and the stained DNA is smearingafter Kpn I digestion. M is the DNA size markers.
Mentions: Plaque purification of the ORFV: With continued viral passages, theinfected primary goat testis cells began to show the cytopathic effect (CPE) and form aviral plaque. The CPE in primary goat testis cells was local and limited on the area ofaffected cells after four to five days after infection (Fig. 1AFig. 1.

Bottom Line: ORFV stimulated human monocytes (THP-1) secreting proinflammatory cytokines IL-8 and TNF-α.Similarly, mice infected with ORFV via both intramuscular and subcutaneous routes at two days prior to IAV infection significantly decreased the replication of IAV.In summary, the results of a current study indicated our Hoping strain harbors the immune modulator property; with such a bio-adjuvanticity, we further proved that pre-exposure of ORFV protects animals from subsequent IAV infection.

View Article: PubMed Central - PubMed

Affiliation: Department of Veterinary Medicine, College of Veterinary Medicine, National Chung Hsing University, Taichung 402, Taiwan.

ABSTRACT
Orf virus (ORFV), a member of parapoxvirus, is an enveloped virus with genome of double-stranded DNA. ORFV causes contagious pustular dermatitis or contagious ecthyma in sheep and goats worldwide. In general, detection of viral DNA and observing ORFV virion in tissues of afflicted animals are two methods commonly used for diagnosis of orf infection; however, isolation of the ORFV in cell culture using virus-containing tissue as inoculum is known to be difficult. In this work, the ORFV (Hoping strain) isolated in central Taiwan was successfully grown in cell culture. We further examined the biochemical characteristic of our isolate, including viral genotyping, viral mRNA and protein expression. By electron microscopy, one unique form of viral particle from ORFV infected cellular lysate was demonstrated in the negative-stained field. Moreover, immunomodulating and anti-influenza virus properties of this ORFV were investigated. ORFV stimulated human monocytes (THP-1) secreting proinflammatory cytokines IL-8 and TNF-α. And, pre-treatment of ORFV-infected cell medium prevents A549 cells from subsequent type A influenza virus (IAV) infection. Similarly, mice infected with ORFV via both intramuscular and subcutaneous routes at two days prior to IAV infection significantly decreased the replication of IAV. In summary, the results of a current study indicated our Hoping strain harbors the immune modulator property; with such a bio-adjuvanticity, we further proved that pre-exposure of ORFV protects animals from subsequent IAV infection.

No MeSH data available.


Related in: MedlinePlus