Yu Chuan-xin, Zhu Yin-chang, Yin Xu-ren, Ren Jian-gong, Si Jin, Xu Yong-liang, Shen Lin-nan
Jiangsu Institute of Parasitic Diseases, Wuxi 214064.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2002;20(4):201-4.
To observe the protective immunity induced by the nucleic acid vaccine of 21.7 kDa membrane protein molecule of Schistosoma japonicum Chinese mainland strain (SjC 21.7) in BALB/c mice.
A pair of primers (P1 and P2) was synthesized according to the DNA sequence of the SjC21.7. The ORF sequence of SjC21.7 was amplified by PCR, and the Kozark sequence was added to the position of initiator. The gene fragment was inserted into the eukaryotic expression plasmid pcDNA3.1 to form the recombinant plasmid SjC21.7-pcDNA3.1. Forty-eight BALB/c mice were divided into three groups: control, test and boost. Each mouse was injected in quadriceps femoris with plasmid pcDNA3.1 (control) or recombinant plasmid SjC21.7-pcDNA3.1 (test, boost); for the boost group, with additional P35-pcDNA3.1 and P40-pcDNA3.1. All mice were immunized three times with an interval of 2 weeks, challenged each with 45 cercariae of S. japonicum at the 30th day after final immunization. At day 45 after challenge, all mice were sacrificed, the numbers of worms and hepatic eggs were counted. Antibody level in the sera of mice before and two weeks after immunization was determined with ELISA. The expression of the target gene in quadriceps femoris was observed with immunohistochemistry.
The immunohistochemistry analysis showed that there were specific antigens expressed in the local tissue of the test group mice. There was specific IgG in the serum of partial mice in test and boost groups. Compared with the control group, the worm reduction rate was 29.9% and its egg reduction rate 13.8% in the test group; 31.9% and 28.0% respectively in the boost group. The egg reduction rate in the boost group was higher than that of the test group (P < 0.05).
The SjC21.7 nucleic acid vaccine could induce partial protective immunity against Schistosoma japonicum in BALB/c mice.
观察日本血吸虫中国大陆株21.7 kDa膜蛋白分子(SjC 21.7)核酸疫苗诱导BALB/c小鼠产生的保护性免疫。
根据SjC21.7的DNA序列合成一对引物(P1和P2)。通过PCR扩增SjC21.7的开放阅读框序列,并在起始密码子位置添加Kozark序列。将该基因片段插入真核表达质粒pcDNA3.1中,构建重组质粒SjC21.7-pcDNA3.1。48只BALB/c小鼠分为三组:对照组、试验组和加强组。每只小鼠股四头肌注射质粒pcDNA3.1(对照组)或重组质粒SjC21.7-pcDNA3.1(试验组、加强组);加强组额外注射P35-pcDNA3.1和P40-pcDNA3.1。所有小鼠免疫3次,间隔2周,末次免疫后第30天每只小鼠感染45条日本血吸虫尾蚴。攻击感染后第45天,处死所有小鼠,计数虫体和肝脏虫卵数量。用ELISA法检测小鼠免疫前及免疫后2周血清中的抗体水平。用免疫组织化学法观察股四头肌中靶基因的表达。
免疫组织化学分析显示,试验组小鼠局部组织中有特异性抗原表达。试验组和加强组部分小鼠血清中有特异性IgG。与对照组相比,试验组虫体减少率为29.9%,虫卵减少率为13.8%;加强组分别为31.9%和28.0%。加强组虫卵减少率高于试验组(P < 0.05)。
SjC21.7核酸疫苗可诱导BALB/c小鼠对日本血吸虫产生部分保护性免疫。