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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

Pre-treatment of ORFV-infected cell medium prevents A549 cells from type A influenzavirus (IAV) infection. The primary goat testis cells were infected with ORFV (Hopingstrain). At two hr post infection (hpi), the unattached virus was removed by washingwith PBS. The cells were maintained in complete RPMI 1640 medium with 10% FBS. At 6,12 and 24 hpi, the medium was collected and used for treatment of human A549 cells.After 24 hr treatment, the A549 cells were infected with 1 MOI of IAV (PR8 strain). At12 hpi, expression of viral nucleoprotein (NP) of the IAV was analyzed byimmunoblotting (panel A), and the quantitative analysis of NP production was shown inpanel B. The column of each group was the mean (+/–SD) of three independentexperiments. The results were analyzed by the T-test. The P value<0.05 (shown with a star symbol) indicates the difference between 2 groups isstatistically significant.
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fig_004: Pre-treatment of ORFV-infected cell medium prevents A549 cells from type A influenzavirus (IAV) infection. The primary goat testis cells were infected with ORFV (Hopingstrain). At two hr post infection (hpi), the unattached virus was removed by washingwith PBS. The cells were maintained in complete RPMI 1640 medium with 10% FBS. At 6,12 and 24 hpi, the medium was collected and used for treatment of human A549 cells.After 24 hr treatment, the A549 cells were infected with 1 MOI of IAV (PR8 strain). At12 hpi, expression of viral nucleoprotein (NP) of the IAV was analyzed byimmunoblotting (panel A), and the quantitative analysis of NP production was shown inpanel B. The column of each group was the mean (+/–SD) of three independentexperiments. The results were analyzed by the T-test. The P value<0.05 (shown with a star symbol) indicates the difference between 2 groups isstatistically significant.

Mentions: ORFV inhibited influenza virus replication in A549 cells: As an immunemodulator, ORFV has been shown to act as an inhibitor to prevent other virus infection andalso to work as a tumor killer [3, 16, 19, 31]. The primary goat fibroblast cells were infected with1 MOI of ORFV. The ORFV infected cell medium, of which the infectivity of ORFV was underdetection, was collected at 6, 12 and 24 hpi, and overlaid onto A549 cells. After 24 hrtreatment, the A549 cells were then infected with influenza virus PR8 strain for 12 hr. Asignificant decrease of viral NP protein expression was observed in cells pre-treated withORFV infected cell medium (Fig. 4Fig. 4.


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)

Pre-treatment of ORFV-infected cell medium prevents A549 cells from type A influenzavirus (IAV) infection. The primary goat testis cells were infected with ORFV (Hopingstrain). At two hr post infection (hpi), the unattached virus was removed by washingwith PBS. The cells were maintained in complete RPMI 1640 medium with 10% FBS. At 6,12 and 24 hpi, the medium was collected and used for treatment of human A549 cells.After 24 hr treatment, the A549 cells were infected with 1 MOI of IAV (PR8 strain). At12 hpi, expression of viral nucleoprotein (NP) of the IAV was analyzed byimmunoblotting (panel A), and the quantitative analysis of NP production was shown inpanel B. The column of each group was the mean (+/–SD) of three independentexperiments. The results were analyzed by the T-test. The P value<0.05 (shown with a star symbol) indicates the difference between 2 groups isstatistically significant.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig_004: Pre-treatment of ORFV-infected cell medium prevents A549 cells from type A influenzavirus (IAV) infection. The primary goat testis cells were infected with ORFV (Hopingstrain). At two hr post infection (hpi), the unattached virus was removed by washingwith PBS. The cells were maintained in complete RPMI 1640 medium with 10% FBS. At 6,12 and 24 hpi, the medium was collected and used for treatment of human A549 cells.After 24 hr treatment, the A549 cells were infected with 1 MOI of IAV (PR8 strain). At12 hpi, expression of viral nucleoprotein (NP) of the IAV was analyzed byimmunoblotting (panel A), and the quantitative analysis of NP production was shown inpanel B. The column of each group was the mean (+/–SD) of three independentexperiments. The results were analyzed by the T-test. The P value<0.05 (shown with a star symbol) indicates the difference between 2 groups isstatistically significant.
Mentions: ORFV inhibited influenza virus replication in A549 cells: As an immunemodulator, ORFV has been shown to act as an inhibitor to prevent other virus infection andalso to work as a tumor killer [3, 16, 19, 31]. The primary goat fibroblast cells were infected with1 MOI of ORFV. The ORFV infected cell medium, of which the infectivity of ORFV was underdetection, was collected at 6, 12 and 24 hpi, and overlaid onto A549 cells. After 24 hrtreatment, the A549 cells were then infected with influenza virus PR8 strain for 12 hr. Asignificant decrease of viral NP protein expression was observed in cells pre-treated withORFV infected cell medium (Fig. 4Fig. 4.

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