【作者】 刘文超；

【导师】 郑世民；

【作者基本信息】 东北农业大学 ， 基础兽医学， 2016， 硕士

【摘要】 禽网状内皮组织增生病(reticuloendotheliosis,RE)是由禽网状内皮组织增生病病毒(retieuloendotheliosis virus,REV)引起的以免疫抑制为主的肿瘤性疾病。本试验以1日龄SPF雏鸡为研究对象,通过瑞氏染色法、酯酶染色法、MTT法、ELISA、荧光定量PCR技术,较全面系统的检测了雏鸡外周血白细胞,异嗜性粒细胞数量、T淋巴细胞及其对Con A的增殖功能、B淋巴细胞及其对LPS的增殖功能、免疫球蛋白(Ig G、IgA、Ig M)含量及Cyclin D1和p27 mRNA表达的动态变化,旨在进一步揭示REV感染SPF雏鸡后,其对机体外周血免疫能力与Cyclin D1和p27 mRNA表达变化的影响,为更好地防治RE提供重要的科学依据。主要研究结果如下:(1)1日龄SPF雏鸡感染REV后,其外周血白细胞总数、异嗜性粒细胞、T淋巴细胞数量及其对Con A的增殖功能在REV感染后1~42天,均不同程度低于相应对照雏鸡;其中,异嗜性细胞和TANAE+细胞数量均在病毒感染后14~42天,极显著(P<0.01)低于对照雏鸡;而白细胞总数在REV感染后7~42d,与对照雏鸡相比极显著(P<0.01)下降;T淋巴细胞对Con A的增殖能力,在病毒感染后14~21d和42d,与对照雏鸡相比显著下降(P<0.01或P<0.05)。表明SPF雏鸡感染REV后,外周血细胞免疫能力明显受损。(2)1日龄SPF雏鸡感染REV后,其外周血B淋巴细胞对LPS的增殖能力和免疫球蛋白IgG、Ig A、Ig M含量在REV感染后1~42d,均不同程度低于相应对照雏鸡;其中,Ig G、Ig A、Ig M含量在REV感染后14~28天、21~42d和42d,均显著(P<0.05)或极显著(P<0.01)低于相应对照雏鸡。表明SPF雏鸡感染REV后,外周血体液免疫能力受到严重抑制。(3)1日龄SPF雏鸡感染REV后,其外周血Cyclin D1和p27 m RNA表达在病毒感染后7~42d,均高于相应对照雏鸡,其中Cycling D1 mRNA表达极显著(P<0.01)高于相应对照组;表明Cyclin D1与p27 m RNA过表达和REV所致的细胞增殖密切相关。更多还原

【Abstract】 Reticuloendotheliosis(RE) was an immune suppression of tumor disease of chicks by retieuloendotheliosis virus(REV). One day old SPF chicks were used in this study. Wright’s stain, Esterase staining, the technologies of MTT, ELISA, realtime fluorescence quantitative PCR were used for revealing the number of white cell, heterophilic granulocytes, TANAE+ cell, the content of Ig S(IgA, Ig G, Ig M), the proliferative function of T and B lymphocytes, the dynamic change of Cyclin D1, p27 mRNA expression. Aimed at further revealed the influence of immunity ability and Cyclin D1, p27 mRNA expression in peripheral blood of chicks infected with REV. To provide important scientific basis for prevention and treatment of RE. The results were as follows:(1)One day old SPF chicks infected with REV, the number of white cells,heterophilic granulocytes, TANAE+ and the proliferative function of T lymphocytes in peripheral blood of chicks were lower than that of control group at 1~42 days; the number of heterophilic granulocytes, TANAE+ of infected chicks were significantly lower than that of control group at 14~42 days(P<0.01); the number of white blood cells of infected chicks were significantly lower than that of control group at 7~42 days s(P<0.01); the proliferative function of T lymphocytes in peripheral blood of chicks were significantly lower than that of control group at 14~21 days and 42 day(P<0.01 or P<0.05). These result indicated that the function of cellular immunity in peripheral blood of chickens infected with REV was reduced.(2)One day old SPF chicks infected with REV, the proliferative function of B lymphocytes and the content of Ig A, Ig G, Ig M in peripheral blood of chicks were lower than that of control group at 1~42 days; the content of Ig A, IgG, Ig M in peripheral blood of chicks were lower than that of control group at 21~42 days, 14~28 days, 42 days(P<0.01 or P<0.05); These result indicated that the function of humoral immunity in peripheral blood of chickens infected with REV was reduced.(3)One day old SPF chicks infected with REV, the expression of Cyclin D1, p27 mRNA in peripheral blood of chicks were higher than that of control group at 7~42 days, the expression of Cyclin D1 mRNA were higher than that of control group(P<0.01). These result indicated that the overexpression of Cyclin D1, p27 mRNA wereclosely related with cell proliferation by virus infected.更多还原