猪接触性胸膜肺炎分子细胞病理学及rompP2诱导的保护力

发布时间：2018-12-13 06:48

【摘要】：接触传染性猪胸膜肺炎是由猪胸膜肺炎放线杆菌感染引起的危害严重的急性呼吸道传染性疾病,研究其致病的分子细胞机制和疫苗有重要意义。探讨革兰氏阴性菌胸膜肺炎放线杆菌感染小鼠致肺病变的分子细胞机制。通过滴鼻法感染小鼠,观察肺组织病变。RT-PCR和qPCR方法检测小鼠肺部的Caspase-1等细胞因子表达量的变化。结果发现小鼠在感染后观察到小鼠肺部有严重出血和炎症。实验组小鼠肺组织IL-1β、IL-6、IL-18和TNF-a的表达水平显著增加：TLR4的表达量显著上调；肺部组织Caspase-1的表达水平明显升高,而Caspase-3的表达水平未见明显变化。由此可以得出,TLR-4信号通路可能参与App感染引起的肺部炎症的发生,App感染后激活Toll.-4信号通路使有关细胞释放炎症因子,引起的肺部炎症。炎症可能涉及Caspase-1参与的细胞焦亡。为验证APP外膜蛋白P2是否可能具有成为疫苗的候选分子,诱导小鼠抗攻击感染的保护力。利用基因重组技术制备重组APP外膜蛋白P2 (rompP2), rompP2加铝佐剂皮下注射免疫小鼠,免疫前和感染前ELISA检测小鼠血液抗体水平变化,第3次免疫后2周攻击感染观察小鼠的免疫保护效果。结果表明,rompP2皮下注射免疫可诱导小鼠产生免疫应答和较高的抗攻击感染的免疫保护力。App ompP2的体外重组蛋白rompP2具有很好的免疫原性和一定的诱导抗攻击感染的免疫保护力。
[Abstract]:Exposure to infectious porcine pleuropneumonia is a serious acute respiratory infectious disease caused by Actinobacillus pleuropneumoniae. It is of great significance to study the molecular cellular mechanism and vaccine of infectious porcine pleuropneumonia. To investigate the molecular cellular mechanism of actinobacillus pleuropneumoniae infection in mice with Gram-negative bacteria. RT-PCR and qPCR methods were used to detect the expression of Caspase-1 and other cytokines in the lung of mice. The results showed that severe hemorrhage and inflammation were observed in the lungs of mice after infection. The expression levels of IL-1 尾, IL-6,IL-18 and TNF-a in the lung tissues of the experimental group were significantly increased: the expression of TLR4 was up-regulated, the expression of Caspase-1 in the lung tissue was significantly increased, but the expression level of Caspase-3 was not significantly changed. It can be concluded that the TLR-4 signaling pathway may be involved in the occurrence of pulmonary inflammation induced by App infection, and the activation of Toll.-4 signaling pathway after App infection can release inflammatory factors and cause pulmonary inflammation. Inflammation may involve the involvement of Caspase-1 in the pyrolysis of cells. In order to verify whether APP outer membrane protein P2 may be a candidate molecule for vaccine, it can induce the protection of mice against attack infection. The recombinant APP outer membrane protein P2 (rompP2) was prepared by gene recombination technique. RompP2 and aluminum adjuvant were injected subcutaneously to immunize mice. The changes of blood antibody levels were detected before and before infection with ELISA. The immune protection of mice was observed 2 weeks after the third immunization. The results showed that the subcutaneous injection of rompP2 could induce the immune response and the immune protection of mice against attack infection, and the recombinant protein rompP2 of App ompP2 had good immunogenicity and certain immune protection against attack infection.
【学位授予单位】：湖南农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.28

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