日本血吸虫中国大陆株23kDa膜蛋白核酸疫苗对猪免疫保护作用的研究  

PROTECTIVE EFFECTS OF 23 kDa MEMBRANE PROTEIN DNA VACCINE OF SCHISTOSOMA JAPONICUM CHINESE STRAIN IN INFECTED PIGS

作　　者：朱荫昌[1] 任建功[1] D.A.Harn 徐明[1] 余传信[1] 司进[1] 叶萍[3] 殷旭仁[1] 何伟[1] 许永良[1] 曹国群[1] 华万全1 

机构地区：[1]江苏省血吸虫病防治研究所,无锡214064 [2]美国哈佛大学公共卫生学院 [3]中国农科院上海家畜寄生虫病研究所

出　　处：《中国血吸虫病防治杂志》2002年第1期3-7,共5页Chinese Journal of Schistosomiasis Control

基　　金：联合国发展开发署/世界银行 /世界卫生组织热带病研究与培训特别规划署的资助 ( IDNO.9810 5 1);江苏省卫生厅科教处的部分资助 ( No.H9918)~~

摘　　要：目的　研究日本血吸虫中国大陆株 2 3k Da膜蛋白 (Sj C2 3) DNA疫苗诱导猪的保护性免疫作用。方法　构建并大量制备 pc DNA3.1- Sj C2 3质粒 DNA和 pc DNA3.1- p35 ,pc DNA3.1- p40(小鼠 IL - 12的 2个亚单位 )的 DNA疫苗。 30头 2 .5月龄的上海松江本地猪 (阉猪 )分为 A、B、C3组 ,每组 10头。 A组 (Sj C2 3组 )于每头猪两侧臀部肌肉注射 5 0 0 μg pc DNA3.1- Sj C2 3质粒 DNA,B组 (Sj C2 3+IL- 12组 ) ,于每头猪肌肉注射 5 0 0 μg pc DNA3.1- Sj C2 3DNA及 5 0 0 μg pc DNA3.1- p35和 5 0 0μg pc DNA 3.1- p40的混和质粒 DNA ;C组 (对照组 )每头猪肌肉注射 5 0 0μg pc DNA3.1质粒DNA。共免疫 3次 ,每次间隔 3周。末次免疫后 30 d,每猪经背部皮肤攻击感染日本血吸虫尾蚴 6 0 0条。攻击后 45 d剖杀 ,经胸主动脉灌冲 ,并从门静脉收集计算成虫。从肝脏的同一部位切下小块肝组织 ,置 5 % KOH内消化后 ,计算每克肝组织虫卵数 (EPG)。比较各组间减虫率、减雌率和减卵率。在免疫前 ,末次免疫后和攻击后从猪耳静脉采血 ,分离血清 ,用 EL ISA检测抗 Sj C2 3抗体。结果　Sj C2 3组与 Sj C2 3+IL - 12组与质粒对照组相比 ,分别获得 2 9.2 %和 5 8.6 %的减虫率、5 0 .8%和5 8.8%的减雌率以及 48.2 %和 5 6 .4%Objective To induce protective effects immunized with 23kDa membrane protein DNA vaccine of Schistosoma japonicum Chinese strain in infected pigs. Methods Construction and preparation of pcDNA3.1 SjC23 plasmid DNA and pcDNA3.1 p35, pcDNA3 1 p40 (two subunits of IL 12) plasmid DNA Thirty of Chinese Songjiang native castrated pigs were divided into A,B,C three groups. In group A (Sjc23 group), each pig was immunized with 500 μg of SjC23 plasmid DNA by intramuscle (i.m.) in both buttocks. In group B (SjC23+IL 12 group) each pig was immunized with the mixture of 500 μg of SjC23 plasmid DNA and 500 μg of pcDNA3 1 p35?500 μg of pcDNA3 1 p40 DNA by i.m.. In group C (control group) each pig was immunized with 500 μg of pcDNA3 1 as the control. All were immunized for three times with three weeks interval. At day 30 after last immunization, each pig was challenged with 600 of cercariae of Schistosoma japonicum on the back of pigs. At day 45 the pigs were sacrificed and perfused. The adult worms recovered were counted. Small piece of each pig's liver was cut and digested in 5% KOH, and the eggs were counted. The worm reduction rate, female reduction rate and eggs per gram liver tissue (EPG) reduction rate between groups were compared. The sera before immunization, after last immunization and after challenge were collected from pig's ear vein. The antibody response to SjC23 was detected by ELISA. Results The worm reduction rates in SjC23 group and SjC23+IL 12 group compared with control group were 29 2% and 58 6%, the female worm reduction rates were 50 8% and 58 8%, and the hepatic egg reduction rates were 48 2% and 56 4% respectively The anti SjC23 antibody detection showed that 4 of 10 pigs in SjC23 group could induce the antibody response, as well as 5 of 10 pigs in SjC23+IL 12 group. Conclusion SjC23 DNA vaccine could induce a good protective immunity in infected pigs. The bigger animals such as pig are better for vaccine experiments.

关 键 词：日本血吸虫 中国大陆株 23kDa膜蛋白 核酸疫苗 猪 保护性免疫 

分 类 号：R383.24[医药卫生—医学寄生虫学] R532.21[医药卫生—基础医学]