生物法合成伤寒O-糖蛋白结合疫苗及其免疫原性评估
发布时间：2019-06-24 13:36
【摘要】：伤寒由伤寒沙门氏菌(Salmonella Typhi)引发,至今在发展中国家仍是备受关注的重要公共卫生问题。文章通过敲除伤寒菌脂多糖合成途径中O-抗原连接酶基因,转入含脑膜炎奈瑟球菌(Neisseria meningitidis)蛋白糖基化途径中糖基转移酶的表达载体,以及改构的重组铜绿假单胞菌(Pseudomonas Aeruginosa)外毒素A(r EPAN29)的表达载体,使细胞内能够诱导合成以伤寒O特异性多糖(O-specific polysaccharides,OPS)为目标抗原、以r EPAN29为载体蛋白的伤寒OPS-r EPAN29糖蛋白复合物,并对纯化所得复合物进行了免疫原性评价。ELISA测定血清抗体滴度表明,r EPAN29作为载体蛋白能有效增加糖链的免疫原性,糖蛋白比单独的多糖能诱导产生更好的免疫应答;3次免疫、间隔3周比间隔2周Ig G滴度稍有提高;而免疫过量的糖蛋白,抗O-多糖的血清抗体效价并无提升。文章为生物法制备多糖-蛋白结合疫苗提供了新思路,理论上也适用于其他革兰氏阴性菌的疫苗研发。
[Abstract]:Typhoid fever, caused by Salmonella typhimurium (Salmonella Typhi), is still an important public health problem in developing countries. In this paper, the O-antigen ligase gene in lipopolysaccharide synthesis pathway of typhoid typhimurium was knockout, the glycosyltransferase expression vector containing (Neisseria meningitidis) protein glycosylation pathway of Neisseria meningitis and the modified expression vector of recombinant Pseudomonas aeruginosa (Pseudomonas Aeruginosa) exotoxin A (r EPAN29 were transferred to induce the synthesis of typhoid O-specific polysaccharide (O-specific polysaccharides,OPS) as the target antigen in cells. The immunogenicity of typhoid OPS-r EPAN29 glycoprotein complex with r EPAN29 as carrier protein was evaluated. Elisa showed that r EPAN29 as carrier protein could effectively increase the immunogenicity of sugar chain, and glycoprotein could induce better immune response than polysaccharide alone. The titer of Ig G in 3 weeks was slightly higher than that in 2 weeks, but the titer of serum antibody against O-polysaccharide was not increased in the immunized glycoprotein and anti-O-polysaccharide. This paper provides a new idea for the preparation of polysaccharide-protein binding vaccine by biological method, and is also suitable for the vaccine development of other Gram-negative bacteria in theory.
【作者单位】： 吉首大学生物资源与环境科学学院;军事医学科学院生物工程研究所病原微生物生物安全国家重点实验室;
【基金】：国家自然科学基金项目(编号:81373316)资助
【分类号】：R392
[Abstract]:Typhoid fever, caused by Salmonella typhimurium (Salmonella Typhi), is still an important public health problem in developing countries. In this paper, the O-antigen ligase gene in lipopolysaccharide synthesis pathway of typhoid typhimurium was knockout, the glycosyltransferase expression vector containing (Neisseria meningitidis) protein glycosylation pathway of Neisseria meningitis and the modified expression vector of recombinant Pseudomonas aeruginosa (Pseudomonas Aeruginosa) exotoxin A (r EPAN29 were transferred to induce the synthesis of typhoid O-specific polysaccharide (O-specific polysaccharides,OPS) as the target antigen in cells. The immunogenicity of typhoid OPS-r EPAN29 glycoprotein complex with r EPAN29 as carrier protein was evaluated. Elisa showed that r EPAN29 as carrier protein could effectively increase the immunogenicity of sugar chain, and glycoprotein could induce better immune response than polysaccharide alone. The titer of Ig G in 3 weeks was slightly higher than that in 2 weeks, but the titer of serum antibody against O-polysaccharide was not increased in the immunized glycoprotein and anti-O-polysaccharide. This paper provides a new idea for the preparation of polysaccharide-protein binding vaccine by biological method, and is also suitable for the vaccine development of other Gram-negative bacteria in theory.
【作者单位】： 吉首大学生物资源与环境科学学院;军事医学科学院生物工程研究所病原微生物生物安全国家重点实验室;
【基金】：国家自然科学基金项目(编号:81373316)资助
【分类号】：R392
【参考文献】
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