中文摘要：

本研究以小鼠为实验动物模型,研究支气管败血波氏杆菌的外膜蛋白（OMP）和其中的有效保护抗原成分（OMP68）的免疫原性及免疫保护作用。本研究利用改进的Wooldridge的方法提取了支气管败血波氏杆菌P11和P13的OMP,利用SDS-PAGE比较了2株之间差异,采用Western-blotting进行了分析并确定了P13菌株OMP（P13-OMP）中的有效保护性抗原成分,采用电洗脱方法获的分子量为68kD的P13-OMP有效保护抗原成分（OMP68）,然后制备了油乳剂P13的全菌、P13-OMP和OMP68免疫抗原。抗体动态变化：将清洁级小鼠70只随机分成7组,10只/组,用油乳剂P13的全菌和不同剂量的P13-OMP和OMP68免疫抗原分别免疫,采用间接ELISA检测免疫小鼠的相应抗体水平并分析抗体动态变化;主动免疫保护试验：将清洁级小鼠80只随机分成8组,10只/组,分别采用制备的油乳剂P13-OMP（OMP25μg/只）、OMP68（25μg/只）和P13菌体免疫抗原在免疫10d后,分别用100LD50的P11和P13腹腔攻毒,然后统计保护率并分析免疫原所提供的免疫保护力。免疫P13-OMP和OMP68油乳剂抗原后,7d时,抗体水平开始逐渐上升,42d时,抗体水平分别达到2^15.3和2^14.8,然后逐渐下降,70d时,抗体水平分别达到2^9.1和2^10.2。主动免疫时,P11和P13攻毒的P13-OMP免疫组分别均得到9/10和9/10的保护,OMP68免疫组分别均得到9/10和10/10的保护,而P13全菌免疫组分别得到4/10和6/10保护,对照组全部死亡。P13-OMP和OMP68抗原均具有良好的免疫原性和免疫保护作用,为支气管败血波氏杆菌OMP亚单位疫苗的研制奠定了良好的理论基础。

英文摘要：

In order to evaluate the immunogenicity and their protection of outer membrane protein of Bordetella bronchiseptica and effective antibody （OMP68, we established a mice model. The research improved on the method of Wooldridge in order to extract OMP of Bordetella bronchiseptica （P13, P11）, and the differences of OMP were analyzed by SDS-PAGE. The P13-OMP was analyzed with Western blotting and OMP68 was gained by the way of electrophoretic elution. Then we maked the effective antibodies of P13-OME Then P13-OMP, OMP68 and whole-cell bacterin （WCB）immunizing antigen was prepared 70 mice were equally assigned to 7 groups at random. They were immunized by P13-OMP, OMP68 and P13-WCB of different dose respectively. The indirect ELISA was used to detect antibody responses. 80 mice were equally assigned to 8 groups at random. Group a and b were immunized by P13-OMP （25 μg） respectively. Groups c and d were immunized by OMP68（25 μg）; Groups e and f were immunized by WCB（P13）. Ten days after the immunizations, group a, c, e and f were challenged with P13. The other groups were challenged with Pll. Then we registered the rate of protective potency and evaluated protection given by P13-OMP, OMP68 and P13-WCB. Results showed that at 42th day, the titer of serum reach the top： P13-OMP（2^15.3）〉 OMP68（2^14.8）. Then, the titer of serum started the slow-moving descent At 70th day, the titer of serum reach the OMP68 （2^10.2）〉 P13-OMP （2^9.1）. P13-OMP and OMP68 group challenged with P13 and Pll can be efectivly protected; P13-WCB group challenged with P13 and P11 can not be efectivly protected; the control group were died out. The P13-OMP and OMP68 of Bordetella bronchiseptica has good immunogenicity and protection, so the results of this study lay good theoretical foundation for OMP subunit vaccine.