波氏杆菌（Bb）是家兔呼吸道传染病重要病原之一，对养兔业危害严重。目前波氏杆菌疫苗免疫效果不十分理想，寻找安全高效疫苗佐剂将为规模兔场波氏杆菌引起的呼吸道感染防治提供可靠保障。本申请项目拟用ECMS-oil(木鳖子皂苷-白油)作为Bb灭活疫苗佐剂，利用酶联免疫吸附试验、淋巴细胞转化试验、Luminex液相芯片技术、荧光定量PCR、流式细胞术等技术，通过对ECMS-oil-Bb 免疫后总抗体及抗体亚类测定、2免后4周免疫兔淋巴细胞细胞因子和淋巴细胞中转录因子等表达变化测定；通过ECMS-oil对于兔天然免疫系统的免疫调节研究，包括研究在不同时间点ECMS-oil在局部注射后对注射局部微环境细胞因子或趋化因子产生影响，注射局部及邻近淋巴结组织免疫细胞动力学变化及功能状态变化的研究以及ECMS-oil对于兔腹腔巨噬细胞免疫调节作用的影响、ECMS-oil体外对兔脾淋巴细胞增殖和相关细胞因子转录因子mRNA的表达测定等内容阐明ECMS-oil对于Bb疫苗的佐剂作用及作用机理。

Rabbit Bordetella bronchiseptica（Bb） is one of important respiratory infectious diseases in rabbit. The disease induced huge economic losses. The immune effects of present bordetella bronchiseptica vaccine were not ideal. In order to find a safe and effective vaccine adjuvant, ECMS-oil（extraction of Cochinchina momordica seed-oil） will be studied for its adjuvant activity on immune responses against Bordetella bronchiseptica. Total and subclass of antibody in rabbit serum against Bb after immunization will be measured by ELISA methods. Concentrations of cytokines from lymphocytes supernatant, the expression of cytokines or transcription factors in splenocytes 4 weeks after 2nd immunization will be measured by Luminex liquid phase chips and fluorescence quantitative PCR respectively. The influence of ECMS-oil on cytokines or chemokines production，kinetics and functional activity of lymphocytes at injection location or neighborhood lymph nodes after several inject time points will be measured by Luminex liquid phase chips method and flow cytometry respectively. The influences of ECMS-oil on phagocytosis activity from peritoneal macrophages, influences of ECMS-oil on the proliferation of lymphocytes and expression of related mRNA of cytokines or chemokines are also going to be measured. These results from the research will illuminate mechanism of the newly and efficient adjuvant.