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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

Detection of the viral B2L gene expression and viral OV20.0 protein in infectedprimary goat cells. (A) RNA was extracted from cells infected with ORFV at 0, 1, 2, 3,12, 20 and 24 hpi (lanes 1–7) for reverse transcription (RT) − PCR detection. Thetranscription of B2L appeared at 2 and 3 hr post-infection and then was salientlyincreased at 12, 20 and 24 hpi. The RT was conducted without reverse transcriptase(−). + indicates a positive control in which DNA template is derived from viruslysate. (B) The expression of OV20.0 (the ortholog of vaccinia virus E3 protein) wasobserved at 6 hr, 12 hr and 24 hr after infection; the expected molecular weight ofOV20.0 is 25 kDa indicated by the arrowhead. Lane 1 is mock infection. Electronmicrograph of orf viruses (C) prepared from primary goat testis cells. Thecharacteristic morphology of orf virus was observed, and viral particles showedovoid-shape with a spiral crisscross pattern (bar=100 nm).
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fig_002: Detection of the viral B2L gene expression and viral OV20.0 protein in infectedprimary goat cells. (A) RNA was extracted from cells infected with ORFV at 0, 1, 2, 3,12, 20 and 24 hpi (lanes 1–7) for reverse transcription (RT) − PCR detection. Thetranscription of B2L appeared at 2 and 3 hr post-infection and then was salientlyincreased at 12, 20 and 24 hpi. The RT was conducted without reverse transcriptase(−). + indicates a positive control in which DNA template is derived from viruslysate. (B) The expression of OV20.0 (the ortholog of vaccinia virus E3 protein) wasobserved at 6 hr, 12 hr and 24 hr after infection; the expected molecular weight ofOV20.0 is 25 kDa indicated by the arrowhead. Lane 1 is mock infection. Electronmicrograph of orf viruses (C) prepared from primary goat testis cells. Thecharacteristic morphology of orf virus was observed, and viral particles showedovoid-shape with a spiral crisscross pattern (bar=100 nm).

Mentions: Viral gene expression examined by RT-PCR and immunoblotting: To examineviral gene expression in the primary goat testis cell, the viral RNA was detected by RT-PCR.Despite the weaker expression, the transcripts of B2L can be detected at the early stage ofinfection (2–3 hpi), and it was largely synthesized after 12 hpi (Fig. 2AFig. 2.


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)

Detection of the viral B2L gene expression and viral OV20.0 protein in infectedprimary goat cells. (A) RNA was extracted from cells infected with ORFV at 0, 1, 2, 3,12, 20 and 24 hpi (lanes 1–7) for reverse transcription (RT) − PCR detection. Thetranscription of B2L appeared at 2 and 3 hr post-infection and then was salientlyincreased at 12, 20 and 24 hpi. The RT was conducted without reverse transcriptase(−). + indicates a positive control in which DNA template is derived from viruslysate. (B) The expression of OV20.0 (the ortholog of vaccinia virus E3 protein) wasobserved at 6 hr, 12 hr and 24 hr after infection; the expected molecular weight ofOV20.0 is 25 kDa indicated by the arrowhead. Lane 1 is mock infection. Electronmicrograph of orf viruses (C) prepared from primary goat testis cells. Thecharacteristic morphology of orf virus was observed, and viral particles showedovoid-shape with a spiral crisscross pattern (bar=100 nm).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig_002: Detection of the viral B2L gene expression and viral OV20.0 protein in infectedprimary goat cells. (A) RNA was extracted from cells infected with ORFV at 0, 1, 2, 3,12, 20 and 24 hpi (lanes 1–7) for reverse transcription (RT) − PCR detection. Thetranscription of B2L appeared at 2 and 3 hr post-infection and then was salientlyincreased at 12, 20 and 24 hpi. The RT was conducted without reverse transcriptase(−). + indicates a positive control in which DNA template is derived from viruslysate. (B) The expression of OV20.0 (the ortholog of vaccinia virus E3 protein) wasobserved at 6 hr, 12 hr and 24 hr after infection; the expected molecular weight ofOV20.0 is 25 kDa indicated by the arrowhead. Lane 1 is mock infection. Electronmicrograph of orf viruses (C) prepared from primary goat testis cells. Thecharacteristic morphology of orf virus was observed, and viral particles showedovoid-shape with a spiral crisscross pattern (bar=100 nm).
Mentions: Viral gene expression examined by RT-PCR and immunoblotting: To examineviral gene expression in the primary goat testis cell, the viral RNA was detected by RT-PCR.Despite the weaker expression, the transcripts of B2L can be detected at the early stage ofinfection (2–3 hpi), and it was largely synthesized after 12 hpi (Fig. 2AFig. 2.

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