肺炎球菌多糖蛋白结合疫苗的研制及其在幼小鼠体内诱导的免疫应答  

Preparation of Pnemnococcal Conjugate Vaccine and Its Inmnmogenicity in Infant Mice

作　　者：陈泽宇[1] 陈兵[1] 王正敏[1] 李忠明 宋丹 迟放鲁[1] 

机构地区：[1]复旦大学附属眼耳鼻喉科医院耳鼻喉科,上海200031 [2]海规生物科技(上海)有限公司,上海200436

出　　处：《中国眼耳鼻喉科杂志》2005年第5期278-280,共3页Chinese Journal of Ophthalmology and Otorhinolaryngology

基　　金：国家自然科学基金资助项目(30471872)

摘　　要：目的研制肺炎球菌多糖蛋白结合疫苗,并通过幼小鼠的动物实验检测其免疫原性。方法用19F 型肺炎球菌多糖以氨基还原法分别与牛血清白蛋白及鸡卵清蛋白偶联结合,以凝胶柱纯化,加入铝佐剂制备成疫苗,然后分别以3种不同多糖含量的结合疫苗腹腔接种3wk 龄幼小鼠,每隔2wk 复种一次,接种3次后采血。多糖对照组幼小鼠以19F 型肺炎球菌多糖疫苗接种,阴性对照组以0.15 M PBS 接种。用 ELISA 法检测动物血清中抗肺炎球菌多糖抗体水平。结果经层析后肺炎球菌多糖蛋白结合物的 OD 峰值较多糖及载体蛋白的各自峰值均明显前移,以多糖含量分别为0.25μg、1.0μg、4.0μg的结合疫苗接种的3组幼小鼠血清中抗肺炎球菌多糖抗体平均滴度分别为1:400、1:800、1:1600,多糖对照组为1:50(P<0.05)。结论已建立制备肺炎球菌多糖蛋白结合疫苗的方法,该疫苗能在幼小鼠体内引起明显的免疫应答。Purpose To preparare the pneumococcal conjugate vaccine and evaluate its immunogenicity in infant mice. Methods Type 19F polysaccharide（PS） was covalently linked to protein carrier bovine serum albumin （BSA） or chicken egg albumin （CEA） by a modified reductive amination method. The conjugate was purified by sepharose CL-4B. The infant mice were immunized with the conjugate vaccine of variate PS dose. The control group （ 19F PS） was immunized with type 19F PS vaccine, The non-immunized control group was injected with 0.15M PBS. The antibody titers to PS in mice serum were measured by ELISA. Results The peak of OD reading of the conjugate shifts remarkably forward compared with that of PS or the proteins. The antibody titer gradually improved as the PS dose increased. The PS （19F） control group has almost not IgG to PS compared with the study group （ P 〈 0.05 ）. Conclusion The procedures to prepare pneumococcal conjugate vaccine have been successfully established. The conjugate vaccine has effective immunogenicity in infant mice rather than PS vaccine.

关 键 词：蛋白结合疫苗 肺炎球菌 中耳炎 免疫预防 多糖蛋白结合疫苗 肺炎球菌多糖 免疫应答 体内诱导 幼小鼠 研制 

分 类 号：R392[医药卫生—免疫学] R764.21[医药卫生—基础医学]