日本血吸虫pcDNA3/SjCWL01核酸疫苗的构建及其对小鼠免疫效果观察  

Construction of the nucleic acid vaccine pcDNA3/SjCWL01 against Schistosoma japonicum and evaluation of its immuno-protective effect on mice

作　　者：彭先楚[1] 汪世平[1] 何卓[1] 吕志跃[1] 李庆华[1] 周松华[1] 李文凯[1] 徐绍锐[1] 

机构地区：[1]中南大学湘雅医学院寄生虫学系,长沙410078

出　　处：《中国人兽共患病学报》2007年第8期752-756,共5页Chinese Journal of Zoonoses

基　　金：国家863重大专项(No.2004AA2Z3530);湖南省"十一.五"重大专项(No.2006SK1001);湖南省"十一.五"重点学科建设经费联合资助

摘　　要：目的构建日本血吸虫pcDNA3/SjCWL01核酸疫苗并进行免疫保护效果观察,评价其作为疫苗的潜能。方法将日本血吸虫基因SjCWL01亚克隆入真核表达载体pcDNA3,构建目的基因真核表达质粒,将pcDNA3/SjCWL01质粒转化大肠杆菌DH5α,大量制备DNA疫苗并免疫小鼠3次,间隔2w,末次免疫后2w,用ELISA法检测免疫鼠血清特异性抗体效价,日本血吸虫尾蚴进行腹部皮肤攻击感染,感染后45d剖杀冲虫,分别计算减虫率,每克肝、粪卵减少率。结果重组DNA疫苗(pcDNA3/SjCWL01)与对照组比较,虫荷、每克肝卵、每克粪卵数分别下降了27.6%,39.5%,45.9%。结论表明pcD-NA3/SjCWL01疫苗可诱导小鼠产生部分抗血吸虫感染的保护力。To evaluate the potential value of the novel gene of S. japonicum SjCWL01 as a nucleic acid candidate vaccine by the construction of pcDNA3/SjCWL01 followed by observation of its immunological protection effect on mice. A novel gene of S. japonicum Sj CWL01 was subcloned into the eukaryotic expression vector pcDNA3. The positive recombinants were identified by PCR and restriction enzyme digestion. Preparation of the DNA vaccine was made by transforming the plasmid pcDNA3/SjCWL01 into E. coli DHSa. The experimental mice were immunized with purified pcDNA3/SjCWL01 DNA vaccine and challenged with 40-t-1 S. japonicum cercariae percutaneously at the 2nd week after the third immunization. Levels of the specific antibody were detected by ELISA before infection. The reduction rates of the worm burden, liver eggs per gram and fe- cal eggs per gram were examined 45 days after challenge infection. Compared with the control group, the reduction rates of the worm burden, liver eggs per gram and fecal eggs per gram were 27.6%, 39.5% and 45.9% in the experimental mice, respectively. The pcDNA3/SjCWL01 nucleic acid vaccine can induce partial protective immunity against S. japonicum infection in mice.

关 键 词：日本血吸虫 SjCWL01 DNA疫苗 免疫保护 

分 类 号：R383[医药卫生—医学寄生虫学]