| Reticuloendotheliosis(RE)is one of the important immunosuppressive neoplastic diseases of poultry,but because of its clinical symptoms are not typical,the disease has not been given enough attention for a long time.Nowadays,Reticuloendotheliosis(RE)prevention and control has been gradually attached importance on poultry industry in China.The disease caused by reticuloendotheliosis virus(REV),the biological characteristics of immune suppression,viral,tumorigenicity.Cyclin D1 is a family of proteins closely related to cell cycle function,by binding and activation to a specific protein kinases and activating its activity,it played a key role in regulating the cell cycle G1 to S phase.1-day-old SPF chickens were used in this study.On the basis of successful preparation of rabbit polyclonal antibody against chicken Cyclin D1,The cell cycle distribution,the number of and the ratio of CD4+ and CD8+T of lymphocytes in immune organs was detected by flow cytometry;Cell culture combined with MTT assay were used to detect the changes of proliferative function of T and B lymphocytes in immune organs;The expression of Cyclin D1 mRNA in immune organs was detected by Real Time-PCR,and the expression of Cyclin D1 protein was detected by indirect ELISA.The effects of REV infection on the immune function and the expression of Cyclin D1 in immune organs of SPF chickens were studied comprehensively.To provide the basis for prevention and treatment of RE.The results were as follows:(1)The proliferative function of T lymphocytes in the thymus and spleens of 1-day-old SPF chickens after infected with REV 1-49 days is lower than that in control chickens in varying degrees,both 14-28 days were significantly lower than control chickens after infected with REV(P<0.01);there were no statistical differences in bursa of Fabricius;the number of CD4+T lymphocytes in the thymus was significantly lower than that in the control chickens at 14-28 days after infected with REV(P<0.05 or P<0.01),the number of CD8+T lymphocytes was extremely significantly lower than that in control chickens at 21-35 days after infected with REV(P<0.01);The number of CD4+T lymphocytes in the spleens were extremely significant lower than that in the control chickens(P<0.05 or P<0.01)at the 7-35 days after infected with REV,Compared with the control chickens,the number of CD8+T lymphocytes was significantly decreased(P<0.01)on the3 rd day,14 th day and 28 th day after REV infection.The number of CD4+T lymphocytes in the bursa of Fabricius was significantly decreased than that in the control chickens at 3-14 days and 28 days after REV infection(P<0.01),compared with the control chickens,the number of CD8+T lymphocytes was significantly lower than that of the control chickens at fist day after REV infection(P<0.01),and significantly higher than that in the control chickens at the 7th to 49 th daysafter infected with REV(P<0.01 or P<0.05).Compared with the control chickens,the ratio of CD4+/CD8+T lymphocytes in thymus,spleens and bursa of Fabricius were decreased in varying degrees after REV infection(P<0.05 or P<0.01).The results showed that the immune function of the main immune organs(thymus,spleens and bursa of Fabricius)of 1 day-old SPF chickens was damaged in varying degrees after infected with REV.(2)The proliferative function of B lymphocytes in the bursa of Fabricius of 1-day-old SPF chickens after infected with REV 14-35 days is significantly lower than that in control chickens(P<0.01),the remaining difference was not statistically significant(P>0.05).In the spleens,compared with the control chickens,the proliferative function of B lymphocytes was significantly decreased(P<0.01)on the 14-49 days after infected with REV(P<0.05)The remaining difference was not statistically significant(P>0.05).The results showed that the humoral immune function of the bursa of Fabricius was decreased in varying degrees of decline.(3)In the cell cycle,the G0/G1 and S period of lymphocytes in the thymus,spleens and the bursa of Fabricius of 1-day-old SPF chickens after infected with REV 14-28 days is higher than that in control chickens in varying degrees.The cells were mainly concentrated in the S phase for DNA replication,G2 / M phase cells were relatively few.A large number of cells stagnated in the S phase.Indicating that cell proliferation was inhibited.(4)The expression of Cyclin D1 mRNA in the thymus of 1-day-old SPF chickens infected with REV was higher than that of the control chickens in varying degrees.Among them,14-28 days after infected with REV,the expression of Cyclin D1 mRNA in the thymus was significant higher than that control chickens(P<0.05).Compared with the control chickens,the expression of Cyclin D1 protein was significantly decreased(P<0.01 or P<0.05)at 7-14 days after REV infection.The expression of Cyclin D1 m RNA in the spleens was significantly higher than that of the control chickens on the 14-28 th day after REV infection(P<0.05).There was no statistically significant difference(P>0.05).Compared with the control chickens,the expression of Cyclin D1 protein in the spleens was significantly increased(P<0.05 or P<0.01)at 1-49 days after infected with REV.The expression of Cyclin D1 mRNA in bursa of Fabricius was significantly higher than that in control chickens at 21-28 days after infected with REV(P<0.05 or P<0.01).Compared with the control chickens,the expression of Cyclin D1 protein in bursa of Fabricius was significantly increased on the 14 th day to 49 th day after REV infection(P<0.05 or P<0.01).The results showed that the expression of Cyclin D1 mRNA and the expression of Cyclin D1 protein in immune organs of SPF chickens were closely related to the immune suppression and cell cycle distribution induced by REV. |
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