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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

Expression of IL-8 and TNF-α in ORFV treated human monocyte THP-1 cells. To examinewhether our ORFV can affect the cellular cytokine levels, THP1 cells were incubatedwith 10 MOI of ORFV for 48 hr. The cell medium was collected for examination of thecytokine IL-8 and TNF-α expressions by using the ELISA kit (KOMA BIOTECH INC.) Theresults demonstrated that IL-8 and TNF-α expression level significantly increased incells infected with ORFV, compared with that of PBS control. The column of each groupwas the mean (+/–SD) of three independent experiments. Statistical analysis wasperformed using unpaired T-test, and P value <0.05 (shown with astar symbol) indicates the statistical significance.
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fig_003: Expression of IL-8 and TNF-α in ORFV treated human monocyte THP-1 cells. To examinewhether our ORFV can affect the cellular cytokine levels, THP1 cells were incubatedwith 10 MOI of ORFV for 48 hr. The cell medium was collected for examination of thecytokine IL-8 and TNF-α expressions by using the ELISA kit (KOMA BIOTECH INC.) Theresults demonstrated that IL-8 and TNF-α expression level significantly increased incells infected with ORFV, compared with that of PBS control. The column of each groupwas the mean (+/–SD) of three independent experiments. Statistical analysis wasperformed using unpaired T-test, and P value <0.05 (shown with astar symbol) indicates the statistical significance.

Mentions: Detection of cytokines produced in THP-1 cells with ORFV: The ORFVinfection elicits expression of proinflammatory cytokines, such as several interleukins(ILs) and TNF-α, which contributes to the immune regulation of ORFV and has beendemonstrated in many studies [8, 9, 15, 26, 37]. It is important toexplore whether ORFV (Hoping strain) harbors the immunostimulating activity. The humanmonocyte cell line, THP-1 cell, was infected with our local isolate at MOI of 10 for 48 hr.The cell medium was collected for detection of IL-8 and TNF-α cytokine production. As theresult shown in Fig. 3Fig. 3.


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)

Expression of IL-8 and TNF-α in ORFV treated human monocyte THP-1 cells. To examinewhether our ORFV can affect the cellular cytokine levels, THP1 cells were incubatedwith 10 MOI of ORFV for 48 hr. The cell medium was collected for examination of thecytokine IL-8 and TNF-α expressions by using the ELISA kit (KOMA BIOTECH INC.) Theresults demonstrated that IL-8 and TNF-α expression level significantly increased incells infected with ORFV, compared with that of PBS control. The column of each groupwas the mean (+/–SD) of three independent experiments. Statistical analysis wasperformed using unpaired T-test, and P value <0.05 (shown with astar symbol) indicates the statistical significance.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig_003: Expression of IL-8 and TNF-α in ORFV treated human monocyte THP-1 cells. To examinewhether our ORFV can affect the cellular cytokine levels, THP1 cells were incubatedwith 10 MOI of ORFV for 48 hr. The cell medium was collected for examination of thecytokine IL-8 and TNF-α expressions by using the ELISA kit (KOMA BIOTECH INC.) Theresults demonstrated that IL-8 and TNF-α expression level significantly increased incells infected with ORFV, compared with that of PBS control. The column of each groupwas the mean (+/–SD) of three independent experiments. Statistical analysis wasperformed using unpaired T-test, and P value <0.05 (shown with astar symbol) indicates the statistical significance.
Mentions: Detection of cytokines produced in THP-1 cells with ORFV: The ORFVinfection elicits expression of proinflammatory cytokines, such as several interleukins(ILs) and TNF-α, which contributes to the immune regulation of ORFV and has beendemonstrated in many studies [8, 9, 15, 26, 37]. It is important toexplore whether ORFV (Hoping strain) harbors the immunostimulating activity. The humanmonocyte cell line, THP-1 cell, was infected with our local isolate at MOI of 10 for 48 hr.The cell medium was collected for detection of IL-8 and TNF-α cytokine production. As theresult shown in Fig. 3Fig. 3.

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