Dynamic Of IFN-г,IL-2 And IL-6 Expression In Spleen And Pbmc Of NDV Immunized Chickens

The pathogen of Newcastle disease (ND) , Newcastle disease virus (NDV) , could cause an acute.highly contagious viral disease characterized by breath difficulty,diarrhea,nervous function disturbance and hemorrhagic sites on the serosal and mucosal surface. Newcastle disease virus (NDV) belong to papramyxovirus,papramyxoviridae. It has a wide host range, infecting 27 of the 50 orders of birds.The infectious ratio of non-typical ND was increasing recently and theatened the development of avain farming seriously.Seven pairs of primers were designed according to the published gene sequences of avian IFN- y , IL-2, IL-6, β -actin with Oligo6.1 and Primer 5.0 software.The partial sequence of IFN- y , IL-2, IL-6 were amplified with RT-PCR from the total RNA extracted from spleen and periphera blood mononuclear cell (PBMC),sequencing analysis showed that the results were correct. Thus mRNA of above cytokines could be quantitied and RT-PCR detection technology was established.The detection results of NDV immunized chickens showed that, in comparision with control group, the changes of mRNA expression quantity of IFN- y of V4 total virus and NDV La Sola strain P5 pure peptide immunized groups were small (P>0.05) ;those of the other groups reduced dramasticlly.The experiment results of CpG motif and NDV peptide immunication were follows.The expression level ,in comparision with that of NDV control group ,of three cytokines of Linear CpG immunized group was most high,the effect of immunity enhance was most apparent (P< 0.05) while the increasing of expression level of three cytokines of stem-loop CpG immunized group and the effect of immunity enhance was not apparent (P>0.05). The expression level of IL-6 mRNA of linear CpG immunized group was much more high than other groups and the expression quantity of IL-6 mRNA among three cytokines was most high.In this study, a RT-PCR technology was established to detect mRNA expression quantity of avian cytokines successfully and the dynamic law of IFN- y , IL-2, IL-6 mRNA expression quantity in periphera blood mononuclear cell (PBMC) of NDV immunized chickens were studied. The detection method is easy for estimation and comparative analysis as a good immunity model.In short,the study will be benefit for further study of complex immunication respond after NDV immunication and immunication enhance effect of CpG It provides the basis evidence and material basis for ND prevention.