摘要
应用基因免疫方法制备日本血吸虫Sj22蛋白的抗体,并研究CpG佐剂和蛋白加强策略在增强基因免疫效果中的作用。采用肌肉注射的方法免疫小鼠,实验动物分为4组:A组注射pVAX1-sj22质粒100μg/只;B组注射pVAX1-sj22质粒50μg/只,同时注射CpG佐剂20μg/只;C、D组注射pcDNA3.1-sj22质粒100μg/只。分别在0、3、6周进行免疫,第9周A、B、C组用30μg重组sj22蛋白加强免疫,D组则用pcDNA3.1-sj22质粒100μg加强免疫。第0、9、10周割尾采血,使用ELISA方法进行抗体效价测定,Westernblot验证抗体的特异性。结果表明:使用基因免疫的方法获得了针对Sj22的特异性抗血清,CpG佐剂能够有效降低质粒用量,基因免疫-蛋白加强的策略使得抗体的滴度提高了40～1280倍。
To study the genetic immunization effectiveness, 16 6-week-old female BALB/C mice were immunized with recombinant plasmids with or without CpG adjuvant, and then boosted with the same recombinant plasmids or the recombinant protein of Schistosoma Japonicum antigen Sj22 in four experimental groups. Mice in group A were immunized with 100μg of plasmid pVAX1-sj22. Mice in group B were immunized with 50μg plasmid pVAXl-sj22 and 20μg CpG adjuvant simultaneously. Mice in group C and D were immunized with pcDNA3.1-sj22. All the mice were immunized at the 0, 3rd, and 6th week by i.m. injection and boosted with recombinant Sj22 protein except group D boosted with the plasmid pcDNA3.1-sj22 at the 9th week. Blood was collected at the 0, 9th and 10th week, and the titer of the serum was detected by ELISA, the specificity of the antibody was examined by Western blot. In conclusion, the antiserum specified to Sj22 protein was obtained by genetic immunization. And it showed that the application of CpG adjuvant can reduce the amount of the plasmid and the DNA prime-boost protein strategy can improve the effectiveness of immunization reaction in genetic immunization.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2007年第1期6-10,共5页
China Biotechnology
基金
国家"863"计划资助项目(2004AAZE1010)
国家"973"计划资助项目(2006CB708510)
上海市重大基础项目(04DZ14010)
