| Newcastle disease and avian influenza are still two main diseases in poultry production,with high mortality and huge economic losses to the poultry industry.At present,the two diseases are mainly prevented by vaccines.However,new type viruses appear constantly,and there are some problems,such as poor immune effect,immune tolerance and immune failure,in clinical immunization practices.Therefore,it is of theoretical and practical significances to study the molecular mechanisms of immune response and develop practical and feasible early diagnostic methods for detecting immune effects.In this study,chickens were immunized by Newcastle disease vaccine(?line Lasota strain)and avian influenza vaccine(H9 subtype,SS strain),and the expressional characteristics of immune related genes(akirin2,miR-155 and SMS1)were analyzed in the natural immune stage and specific immune stage after inoculation with vaccines.The main research methods and results were as follows:In order to further study the expression and regulation of immune related genes in chickens immunized with vaccines,the Hi-Line Brown chickens of 11 days old were used as the research models,and were immunized with Newcastle vaccines and avian influenza vaccines respectively.Then,liver,thymus,bursa,spleen and blood were collected at 12h,24h,72h,120h,7d,21d,35d and 42d after immunization,respectively.The transcriptional activities of immune related gene akirin2 were analyzed by fluorescence quantitative PCR.The results showed that the antibody levels of the Newcastle disease vaccine group and the avian influenza vaccine group all reached the peak at 14d.In the Newcastle disease vaccine group,akirin2 gene was down-regulated significantly in liver,thymus and bursa at 72h(P<0.01),but up-regulated significantly in liver,thymus,spleen and bursa at 120h(P<0.01);In addition,akirin2 gene was up-regulated significantly in blood at 120h(P<0.01).In the avian influenza vaccination group,akirin2 gene was up-regulated in several immune organs,and up-regulated significantly in spleen,thymus and bursa at120h(P<0.01);Akirin2 gene was also up-regulated significantly in thymus and liver at 7d(P<0.01).Interestingly,akirin2 gene also increased significantly in blood at 120 h(P<0.01).Chickens were immunized with Newcastle disease vaccine and avian influenza vaccine,and serum was collected at 12h,24h,72h,120h,7d,21d,35d and 42d after immunization.Expressional activities of miR-155 were analyzed by fluorescence quantitative PCR in serum.The results showed that mi R-155 was up-regulated significantly at 120 h in the early stage of specific immune response after the immunization with Newcastle disease vaccine(P<0.01);In the avian influenza vaccine group,miR-155 was also up-regulated significantly to the peak at 120 h after immunization(P<0.01).In order to study the function of chicken SMS1 gene,SMS1 full length coding sequences were cloned from AA~+broilers and Sanhuang broiler,and SMS1 gene from four breeds of chicken(Gallus gallus,Hi-Line Brown layer,AA~+broiler and Sanhuang broiler)were analyzed by bioinformatics firstly.Then,the tissue expressional characteristics of SMS1 gene were analyzed.Finally,the correlations between SMS1 gene and immune response were verified.The results showed that 5 SNPs existed in SMS1 coding sequences from the four breeds chickens.The expressional levels of SMS1 gene from 14-day-old Hi-Line Brown layer and Sanhuang broiler were all the highest in blood,while low expressed in other tissues.In the Newcastle disease vaccine group,SMS1 gene was up-regulated significantly in liver at 120h and 21d(P<0.01),respectively.And SMS1 gene was down-regulated at 120h and up-regulated at 21d in blood;In the avian influenza vaccination group,SMS1 gene was significantly up-regulated in liver at 120h(P<0.01),but down-regulated in liver at 21d(P<0.01).SMS1 gene(P<0.01)at 21d was significantly up-regulated in blood.And SMS1 gene was down-regulated slightly at 120h,but up-regulated significantly at 21d in blood(P<0.05).This study shows that akirin2,miR-155 and SMS1 genes are important regulatory genes in immune responses.Study the expression and regulation of these genes in immune response,can provide an important theoretical references for further understanding the molecular mechanisms of akirin2 gene in the vaccine immune response,the possible application of miR-155 in developing early diagnosis methods of immune effects,and insight into the functional characteristics of SMS1 gene. |
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