福氏2a痢疾杆菌O-特异性多糖与破伤风类毒素结合疫苗的制备及其免疫学特性  

Preparation,characterization and immunogenicity of conjugate vaccine composed of the O specific polysaccharide of S.flexneri 2a and tetanus toxoid

作　　者：谢贵林[1] 冯建军[2] 杜琳[1] 吴应文[2] 

机构地区：[1]兰州生物制品研究所,兰州730046 [2]兰州大学生命科学学院

出　　处：《微生物学免疫学进展》2001年第1期1-7,共7页Progress In Microbiology and Immunology

摘　　要：通过己二酸二肼 (ADH)将福氏 2a痢疾杆菌脂多糖 (LPS)经酸水解脱毒纯化后得到的O SP和破伤风类毒素 (TT)蛋白共价结合 ,制备了福氏 2a痢疾杆菌结合物。以 2 5 μg多糖或含 2 5 μg多糖的结合物经皮下注射免疫NIH品系雌性小鼠 ,同时以 2 5 μg多糖或含 2 5 μg多糖的结合物经耳缘静脉注射家兔。用ELISA方法分别检测小鼠和家兔血清中抗脂多糖抗体水平及抗TT水平 ,并用豚鼠血清补体介导进行了体外杀菌实验。结果显示 :用本实验方法提取的多糖 ,合成的多糖衍生物 ,多糖 蛋白结合物都具有福氏 2a痢疾杆菌O 抗原特异性 ,其化学组成及结构特异性和国外文献报道基本一致。单独用多糖免疫小鼠和家兔未能诱导LPS抗体 ,而结合物免疫小鼠和家兔的血清诱导出了较高的抗LPSIgG抗体及抗TT抗体 ,并有较强的体外杀菌活性。产生的抗体存在着免疫记忆性 。Lipolysaccharide (LPS) is both of the major toxic factor and the major protective antigen of Shigellae.It was considered that O specific polysaccharide (O SP) is the structure of protcetive antigenic.However,O SP alone can not elicit specific IgM and IgG both in human and animals because of its small molecular weight and simple structure which consisted of four repeat saccharide unit.The O SP of Shigella flexneri type 2a was purified and covalently bound to tetanus toxoid(TT) then formed a conjugate O SP TT.2.5μg of O SP or its conjugate was injected subcutaneously into NIH mouse,or 25μg iv injection a rabbit.Serum IgG and IgM antibodies against LPS of S.flexneri type 2a were measured by ELISA,and the bactericidal activity of anti. LPS antibody was measured by complement dependent bactericidal experiments.The results showed that the IgG and IgM antibodies are not detected in the serum of mice and rabbits serum immunized by O SP alone.A statistically significant rise in both of IgM and IgG could be elicited by O SP TT conjugate immunized mice and rabbits,the booster reactions occur after second and third injection as a result of the immunological memory response.Anti LPS antibodies in serum contain the bactericidal activity in confirmation with complement dependent bactericidal test in this study.

关 键 词：福氏2A痢疾杆菌 O-特异性多糖 破伤风类毒素 结合疫苗 免疫原性 

分 类 号：R392-33[医药卫生—免疫学]