日本血吸虫脂肪酸结合蛋白(SjC FABP)核酸疫苗诱导小鼠免疫保护作用研究  

PROTECTIVE IMMUNITY INDUCED BY THE NUCLEIC ACID VACCINE OF FATTY ACIDBIND PROTEIN OF SCHISTOSOMA JAPONICUM CHINESE STRAIN (SJC FABP) IN BALB/C MICE

作　　者：余传信[1] 朱荫昌[1] 殷旭仁[1] 任建功[1] 司进[1] 许永良[1] 沈林南[1] 何伟[1] 

机构地区：[1]江苏省血吸虫病防治研究所,无锡214064

出　　处：《中国血吸虫病防治杂志》2002年第3期163-167,共5页Chinese Journal of Schistosomiasis Control

基　　金：江苏省科技厅应用基础基金 ( BJ980 34 )资助;世界卫生组织 /世界银行 /热带病研究培训特别规划署 ( TDR )项目部分资助

摘　　要：目的　研究日本血吸虫中国大陆株脂肪酸结合蛋白分子 (Sj C FABP)核酸疫苗对 BALB/c小鼠的免疫保护性作用。方法　根据 Sj C2 1.7分子基因序列合成 1对特异性引物 P1和 P2 ,并使其5’端分别带有 Bam H1、Xho1的酶切位点序列 ,采用 PCR方法从重组质粒 Sj C FABP-p UC19中扩增出特异性目的基因的开放阅读框序列 ,并在其翻译起始密码子处引入 Kozark序列。限制性酶切后的基因片段被亚克隆到真核表达质粒 pc DNA3 .1中 ,构建成重组真核表达质粒 Sj C FABP-pc D-NA3 .1,大量制备纯化的重组真核表达质粒。 48只 BAL B/c小鼠分为对照组、实验组、加强组。对照组每只小鼠的股四头肌注射接种 10 0μg质粒 pc DNA3 .1DNA,实验组以同样的方式注射 10 0μg重组质粒 Sj C FABP-pc DNA3 .1DNA,加强组除注射 10 0μg重组质粒 Sj C FABP-PCDNA3 .1DNA外 ,同时注射 P3 5 -pc DNA3 .1,P40 -pc DNA重组质粒各 10 0μg。每隔 2周免疫 1次 ,共免疫3次。第 3次免疫后的第 3 0天每只小鼠经腹部皮肤感染 (4 5± 1)条尾蚴 ,45 d后剖杀 ,计数各组小鼠肝虫卵数及成虫数。于免疫前及末次免疫后 2周分别经尾静脉采血 ,测定抗体水平。在第 2次免疫后第 10天各组取 2只小鼠处死 ,取股四头肌进行免疫组化分析。结果　免疫组化分析显示实?Objective To observe the protective immunity induced by the nucleic acid vaccine offatty acid bind protein of Schistosoma japonicum Chinese strain (SjC FABP) in BALB/c mice. Methods A pair of primers (P1 and P2) was synthesized according to the DNA sequence of the SjCFABP, and the DNA sequences of restrictive enzymes BamH1 and Xho 1 were added to the 5' termi-nator of each primer respectively. The ORF sequence of SjC FABP was amplified by PCR, and the Kozark sequence was introduced at the position of initiator. The gene fragment digested by BamH1 and Xho1 was inserted into the eukaryotic expression plasmid pcDNA3. 1 to form the recombinant plasmid SjC FABP-pcDNA3. 1. The recombinant plasmid was preparated. Forty-eight of BALB/c mice were divided into three groups: control, test and enhanced. Of control group, each mouse was injected with 100 μg plasmid pcDNA3. 1 DNA in quadriceps femoris of hindlimb; of test group,with 100 μg recombinant plasmid SjC FABP-pcDNA3. 1 DNA in quadriceps femoris of hindlimb; of enhanced group, except of 100 μg recombinant plasmid SjC FABP-pcDNA3.1 DNA, each mouse also incubated 100 μg P35-pcDNA3. 1 and 100 μg P40-pcDNA3. 1. All mice were immunized by three times, the interval period was 2 weeks. Each mouse was challenged with 45 cercariae of S.japonicum Chinese strain by abdomen skin at thirtieth day after final immunization. At day 45 after challenge, all mice were sacrificed and perfused, the numbers of recovered worms and hepatic eggs were counted. The antibody level of the sera of mice before immunization and two weeks after immunization was determined with ELISA. At the tenth day after third immunization, two mice of each group were sacrificed and its muscular tissue in quadriceps femoris of hindlimb was analyzed by immunohistochemistry. Results The immunohistochemistry analysis showed that there were specific antigens expressed in the local tissue in the test and enhanced groups. The specific antibody of 8 sera in test group of 13 mice, 6 in enhanced group of 14 mice wer

关 键 词：日本血吸虫 脂肪酸结合蛋白 SjC FABP 核酸疫苗 小鼠 免疫保护 白细胞介素-12 

分 类 号：R532.21[医药卫生—内科学]