| Porcine parvovirus (PPV) is the major causative virus in a syndrome of reproductive failure in swine, which includes stillbirths,mummified fetuses, early embryonic death, and infertility. Moreover, other clinical manifestation besides reproductive failure, such as dermatitis and enteric diseases. Recently, PPV is widespread among swine throughout the world as a signicant economical disease, PPV infection and the harm caused by the upward trend to the pig farming industry and caused great economic losses. as mediate the establishment of an antivirus state and recruit inflammatory cells to the site of infection, Cytokines were clearly not the only factor contributing to disease, And they seemed to be essential for resolution of the infection. The study of pathogenesis of porcine parvovirus were focused on the process of viral replication and infection, while the relationship between cells and virus, variation of cytokines of host cell infected with porcine parvovirus have not been reported. To understand the mechanism of porcine cytokines, clarify the mechanism of PPV, we carried out the following studies:1. The PPV DNA levels and transcript level of cytokines(IFN-β,IFN-γ,IFNAR-1,IFNAR-2,MHC-Ⅰ,MHC-Ⅱ,MX1,iNOS,2-5OAS,RNaseL and IRF-3)of PBMC cultures infected with porcine parvovirus after 1 h, 3 h,6 h, 12 h, 24 h, 48 h, 72 h were detected by quantitative real-time PCR SYBR greenⅠmethod. The virus DNA can be detected an hour after PPV infection, 72 h after infection, the peak value was more then 10 times.The transcript levels of IFNAR-1 were increased significantly on 1 h and 24 h postinoculation(p.i.),especially 1 h p.i the expression levels of IFNAR-1 increased about 89.9 times; IFNAR-2 reached a peak on 24 h p.i, but was not significant at other times;IFN-βreached a peak in 1 h p.i,but decreased on 24 h p.i; The transcript levels of IFN-γwere decreased on 24 h p.i,but significantly increased 48 h p.i; MHC-Ⅰwas increased significantly on 1 h,12 h,48 h,and reached a peak in 48 h; IRF-3,RNaseL were increased significantly on 48 h, but was not significant at other times; MX1,2-5OAS were increased significantly on 24 h, 48 h,and reached a peak in 24 h; MHC-Ⅱwas increased significantly on 1h,3 h,6 h,12 h,and reached a peak in 3 h; PBMC do not secrete iNOS on 1 h p.i,but the transcript levels of iNOS was increased significantly on 3 h p.i, decreased on 24 h p.i. The PPV infection induced PBMC the secretion of anti-virus related cytokines increased significantly.2. The transcript level of cytokines(IL-1β,6,8,12p35,12p40,13,17 and 18)of PBMC cultures infected with porcine parvovirus after 1 h, 3 h, 6 h, 12 h, 24 h, 48 h, 72 h were detected by quantitative real-time PCR SYBR greenⅠmethod. Of the genes levels tested, The transcript levels of IL-1βwere increased significantly on 1h,3 h,6 h,12 h,24 h, the peak value was 58 times on 24 h;IL-6 was increased significantly on 1h,3 h,6 h,12 h,24 h,72 h,and reached a peak on 6 h,which was 40.5 times;IL-12p35 and p40 were decreased significantly on 24 h,but increased significantly on 72 h and 1 h respectively;IL-8 was increased significantly on 1h,3 h,6 h,12 h,24 h,48 h,and reached a peak on 24 h which was 59.7 times;IL-13 was increased significantly on 3 h,but decreased significantly on 12 h,24 h;IL-17 was increased significantly on 1 h,48 h,but decreased significantly on 6 h,12 h,24 h;IL-18 was increased significantly on 6 h. The PPV infection induced PBMC the secretion of inflammatory cytokines increased significantly.3. The transcript level of cytokines(TNF-α,TNFR-2,P53,PBR,Bcl-2,Bcl-xl,Caspase-8 and Fasl) of PBMC cultures infected with porcine parvovirus after 1 h, 3 h, 6 h, 12 h, 24 h, 48 h, 72 h were detected by quantitative real-time PCR SYBR greenⅠmethod. Of the genes levels tested, The transcript levels of TNF-αwere increased significantly on 1 h, then started to decline, decreased significantly on 24 h; TNFR-2 was decreased significantly on 24 h,which was 0.25 times,but increased significantly on 48 h which was 11.5 times;P53 was increased slightly on 1 h and 72 h,but decreased on 6 h,12 h and 24 h;PBR was increased slightly on 1 h,24 h and 48 h, the peak value was 12.9 times on 24 h;Bcl-2 was increased slightly on 1 h,but decreased other times, decreased significantly on 24 h; Bcl-xl was decreased significantly on 24 h,but increased significantly on 72 h; Caspase-8 was increased significantly on 1h and 24 h, the peak value was 19.3 times on 1 h; Fasl was increased significantly on 1h, but decreased significantly on 24 h, which was about 0.02 times. The PPV infection induced PBMC the secretion of apoptosis-related cytokines changed significantly.In summary, the expression of PPV DNA, interferon and related cytokines, inflammatory cytokines,apoptosis-related cytokines of porcine PBMC after infected with PPV were studied. Provide an experiment support for further understand the role of cytokines in the immune mechanism and the molecular mechanisms of PPV, and establish theoretical foundation for screening and development of prevention and treatment of PPV-related drugs. |
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