The Study On Influence Of Transcriptional Profiles Of Porcine Parvovirus And Porcine Circovirus Type2Co-infection On Cytokine MRNA Expression In Peripheral Blood Mononuclear Cells Of Porcine

Porcine parvovirus (PPV) and porcine circovirus type2(PCV2)aretwo viruses which can cause reproductive fail and immunosuppression in infectedpigs．In this study，PPV and PCV2were co-infected on cytokine mRNA expressionin peripheral blood mononuclear cells of porcine. The aim of our study was toexplore the cellular and molecular mechanisms of the effect of PPV and PCV2co-infection on immune functions and Molecular Pathogenesis in porcine.1.The PPV, PCV2DNA levels of PBMCs cultures infected with PPV and/or PCV2co-infected or single after1h,3h,12h,24h,48h,72h were detected byquantitative real-time PCR SYBR greenⅠmethod. The virus DNA can be detectedan hour after PPV, PCV2infection, the DNA content levels of PCV2in PPV andPCV2co-infected higher than PCV2infection. These results indicated that togetherPPV and PCV2co-infection could mutually promote the replication of PCV2.2.The PPV, PCV2DNA levels and transcript level of IL-1β, IL-6, IL-8, IL-12p35,IL-12p40, IL-13, IL-17and IL-18of PBMCs cultures infected with PPV and/orPCV2co-infected or single after1h,3h,12h,24h,48h,72h were detected byquantitative real-time PCR SYBR greenⅠmethod. The results showed that themRNA levels of interleukin cytokines, IL-1β, IL-6, IL-13, IL-8and IL-17cytokineswere significantly increased in co-infected group compared to those of single-infected group in more time points; of them the depression of IL12p35, IL-12p40and IL-18was extremely obvious. These results indicated that the inflammatoryresponse and immunosuppression induced by PPV and PCV2co-infection was moresevere than that of single-infection in the stage of infection．3.The transcript level of IFN-β, IFN-γ, FNAR-1, IFNAR-2, MX1,2-5OAS, RNaseL andIRF-3of PBMCs cultures infected with PPV and/or PCV2co-infected or single after1h,3h,12h,24h,48h,72h were detected by quantitative real-time PCR SYBR greenⅠmethod. The results showed that the mRNA levels of interleukin cytokines,IFN-β, IFN-γ mRNA level were significantly increased in co-infected group comparedto those of single-infected group in more time points, and IFN-γ mRNA level weresignificantly decreased in co-infected group to it of control group; But IFNAR-1mRNAlevel decreased in more time points, and IFNAR-2mRNA level decreased in laterperiod; The IRF-3mRNA level were significantly increased in co-infected groupcompared to those of single-infected group in early period, increased in later period.These results indicated that the mRNA levels of interferon cytokines were low in thewhole test, and activate in early time, inhibition in the later time. Theimmunosuppression induced by PPV and PCV2co-infection was more severe thanthat of single-infection in the stage of infection. The study of effector molecule ofinterleukin MX1,2’5’OAS and RNaseL cytokines show that MX1,2’5’OAS/RNaseL areeffector molecule of interleukin of PPV, and MX1is PPV and PCV2co-infection.4.The transcript level of I MIP-1α, TGF-β1, NF-κB and MURR1of PBMCs culturesinfected with PPV and/or PCV2co-infected or single after1h,3h,12h,24h,48h,72h were detected by quantitative real-time PCR SYBR greenⅠmethod. Of thegenes levels tested, MIP-1α, TGF-β1, NF-κB and MURR1expression was significantlyincreased after virus infected in later period. These results indicated that theimmunosuppression induced by PPV and PCV2co-infection was more severe thanthat of single-infection in the early stage of infection.