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目的 脑膜炎奈瑟菌 (Nm)A群荚膜多糖与B群菌株 34 0 7或 5 42 85 2外膜蛋白复合物偶联 ,希望获得 1种对A和B群Nm感染皆有预防效果 ,并可提高A群荚膜多糖对婴幼儿免疫力的偶联物。方法 用饱和硫酸铵沉淀细菌培养上清液 ,沉淀物再经SephacrylS 30 0纯化获得外膜蛋白复合物。通过碳化二亚铵 (EDC)介导的缩合反应将B群外膜蛋白复合物与A群荚膜多糖偶联。偶联物、未偶联的荚膜多糖、B群外膜蛋白复合物及A群荚膜多糖和B群外膜蛋白复合物简单混合物按相同程序免疫小鼠获得抗体 ,经ELISA、杀菌力试验和Westernblotting测定了偶联物的免疫原性。结果 偶联物比未偶联的荚膜多糖或者荚膜多糖同B群外膜蛋白复合物简单混合抗原的免疫原性增强 2 1~32 0倍 ;其中以B群 34 0 7菌株的外膜蛋白复合物与A群荚膜多糖偶联的效果更好。偶联物免疫血清不仅对A群代表菌株 (2 90 19)和B群菌株 (34 0 7,5 42 85 2和 2 90 2 1)均有较强的杀菌活性 ,而且还对所试的具有不同菌型特征的其它 8株B群菌株仍有较强的交叉反应性。Westernblotting初步分析发现上述两种偶联物免疫的血清与各自的外膜蛋白复合物在相对分子质量约 42× 10 3 、39× 10 3 和 2 6× 10 3 处有相同的反应带 ,其中 42× 10 3 左右的条带为 1类外膜蛋白。结论
Objective To conjugate A group of capsular polysaccharides of Neisseria meningitides (Nm) with the outer membrane protein complex of group B or strain 34 0 7 or 5 42 85 2 in the hope of obtaining a preventive effect on both N and A group Nm infections, And can improve the group A capsular polysaccharide conjugates of infant immunity. Methods The bacterial culture supernatant was precipitated by saturated ammonium sulfate and the precipitate was purified by Sephacryl S 30 to obtain the outer membrane protein complex. Cluster B outer membrane protein complexes were conjugated to Group A capsular polysaccharides by di-carbodiimide (EDC) -mediated condensation reactions. Conjugate, unconjugated capsular polysaccharide, B group of outer membrane protein complex and a simple mixture of group A capsular polysaccharide and B group of outer membrane protein complexes were immunized according to the same procedure to obtain antibodies. ELISA, bactericidal test The immunogenicity of the conjugates was determined by Western blotting. RESULTS: The immunogenicity of the conjugate was 21-130 times higher than that of the unconjugated capsular polysaccharide or the complex mixed antigen of capsular polysaccharide and B outer membrane protein complex. Among them, the outer membrane of 34 0 7 strain of B group Protein complexes and group A capsular polysaccharide conjugate better. The conjugate immune sera not only showed strong bactericidal activity against the representative group A (2 90 19) and group B (34 0 7, 5 42 85 2 and 2 90 2 1), but also tested The other eight strains with different fungal characteristics still had strong cross-reactivity. Westernblotting preliminary analysis found that the serums immunized with the above two conjugates had the same reaction bands with the respective outer membrane protein complexes at about 42 × 10 3, 39 × 10 3 and 26 × 10 3, of which 42 A band of about 10 3 is a type 1 outer membrane protein. in conclusion