不同佐剂对奶牛乳房炎金黄色葡萄球菌多价毒力因子疫苗免疫效果的影响  

The effect of different adjuvants multivalent virulence factor vaccine immunized on mastitis caused by Staphylococcus aureus

作　　者：杨岚[1] 郝永清[1] 潘海婷 何焱[1] 黄海碧[1] 

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018

出　　处：《中国兽医学报》2015年第11期1824-1829,1839,共7页Chinese Journal of Veterinary Science

基　　金：国家科技支撑重点校区项目(2012BAD13B008)

摘　　要：为评价佐剂对金黄色葡萄球菌(S.aureus)CP8-FnBPB-ClfA免疫效果的影响,用纯化的S.aureus血清8型荚膜多糖与FnBPB-ClfA蛋白偶联的偶联物配合不同佐剂对健康成年新西兰大耳白母兔进行免疫接种,第1、2、3、4组的佐剂分别为弗氏佐剂、铝盐佐剂、脂质体佐剂及大肠杆菌不耐热肠毒素B(LTB),第5组为对照组用PBS免疫。用间接ELISA法和Western blot对各组兔血清中的抗体进行监测,评价体液免疫效果;通过检测Th1/Th2类细胞因子含量变化评价机体免疫被激活的程度;T淋巴细胞增殖试验评价细胞免疫,并对三免后2周的白兔进行攻毒保护试验。结果显示,以脂质体为佐剂的组7d时开始产生抗体,为产生抗体最早组,14d后抗体水平就开始下降;以LTB为佐剂的组抗体水平最高,持续时间最长。细胞免疫水平检测结果显示,经ConA刺激后,各组T淋巴细胞普遍增殖,无显著差异,刺激指数都在1.0左右;用抗原刺激的组中,以LTB为佐剂的组T淋巴细胞增殖能力最强,刺激指数为2.23,对白兔的攻毒保护率为80%。The purpose of this study was to assess the ability of adjuvants to enhance Staphylococcus aureus-directed responses following immunization with CP8-FnBPB-ClfA.Healthy adult female New Zealand big ear white rabbits were immunized with either PBS or purified CP8-FnBPBClfA with Freund＇s adjuvant,ALUM,liposome or heat-labile enterotoxin,B subunit fromEscherichia coli（LTB）.Immune reactivity to CP8,FnBPB,ClfA,FnBPB-ClfA and CP8-FnBPB-ClfA was assessed over 60 days by ELISA and western blot.The levels of IL-2,IL-4,IL-10 and IFN-γwere detected by ELISA,the ability of CP8-FnBPB-ClfA to stimulate T cell proliferation was assessed by ex vivo culture and the toxicity test was chosen to detect antibody levels.The adjuvant LTB elicited the best lived CP8-FnBPB-ClfA specific antibody response,closely followed by ALUM and Freund＇s adjuvant.Although the liposome adjuvant group elicited CP8-FnBPB-ClfA antibodies earlier,this response was the shortest lived.Vaccination with ALUM and LTB effectively elicited both Th1 cytokines,IFN-γand IL-2,which participate in cellular immune responses,and the Th2 cytokines,IL-4and IL-10,which participate in humoral.Adjuvant LTB had the strongest multiplication capacity of T lymphocyte.Immunization of rabbits with CP8-FnBPB-ClfA with the LTB adjuvant elicited strong and long lasting of adaptive humoral and cellular immune responses.The survival rate reached 80%.

关 键 词：佐剂 CP8-FnBPB-ClfA 免疫原性 金黄色葡萄球菌 

分 类 号：S852.61[农业科学—基础兽医学]