Construction and Expression of DNA Vaccine pIRES-Sj97-Sj14-Sj26 and Its Immunogenicity in Mice

• Published in: Journal of Huazhong University of Science and Technology. Medical sciences Vol. 27; no. 6; pp. 625 - 629
• Main Author: 刘朔婕 程继忠 唐成武 马彦彬 王淑玉 郭萍 段秋红 高红 戴五星
• Format: Journal Article
• Language: English
• Published: China Department of Biochemistry and Molecular Biology, Tongji Medical College, Huazhong University of Science and Technology,Wuhan 430030 01.12.2007
• Subjects: Animals; Antigens, Helminth - genetics; Antigens, Helminth - immunology; DNA疫苗免疫法; HeLa Cells; Helminth Proteins - biosynthesis; Helminth Proteins - genetics; Helminth Proteins - immunology; Humans; Immunization; Mice; Mice, Inbred BALB C; RNA, Messenger - biosynthesis; RNA, Messenger - genetics; Schistosoma japonicum - genetics; Schistosoma japonicum - immunology; Schistosomiasis japonica - immunology; Schistosomiasis japonica - prevention & control; Vaccines, DNA - biosynthesis; Vaccines, DNA - immunology; 基因结构; 基因表达; 血吸虫病; 预防策略; DNA vaccine immunization; Sj97; Sj26; Sj14; schistosoma japonicum
• ISSN: 1672-0733; 1993-1352
• DOI: 10.1007/s11596-007-0601-4

To find a new preventive strategy for the infection of Schistosoma japonica, plasmid pIRES-Sj97-Sj 14-Sj26 that contains fatty binding protein （Sj 14）, GST （Sj26） and paramyocin （Sj97） that are expressed on the membrane, was constructed. RT-PCR was used to detect the expression of Sj 14 mRNA, Sj26 mRNA and Sj97 mRNA in the Hela cells, the indirect immunofluorescent test was employed for the detection of the expression of trans-membrane Sj26 after the plasmid was transfected into Hela cells. Fifty BALB/c mice were randomly divided into 5 groups and plRES-Sj97-SjI4-Sj26 plasmid DNA, plRES-Sj 14-Sj26 plasmid DNA, plRES-Sj26 plasmid DNA, plRES blank vector and normal saline were respectively injected into the quadriceps muscles of thigh Eight weeks after the immunization the mice were killed and significantly higher level of IgG was detected in the plRES-Sj97-Sj 14-Sj26 group as compared with the plRES blank vector, normal saline and plRES-Sj26 groups （P〈 0.01） and the plRES-Sj 14-Sj26（P〈0.05）. Single splenocyte suspension was prepared to detected the level of IFN-T by ELISA and the lymphocyte stimulating index （SI） by MTT. SI was significantly higher of in the plRES-Sj97-Sj 14-Sj26 group than in other groups （P〈 0.01）, while the IFN-T level was significantly higher the plRES-Sj97-Sj 14-Sj26 group than in plRES blank vector and normal saline groups （P〈0.01）, but no significant differences were found when compared with plRES-Sj 14-Sj26 and plRES-Sj26 groups. Flow cytometery showed that the percentages of CD4＋ and CD8＋ T cells were much higher in the plRES-Sj97-Sj 14-Sj26 group （P〈 0.01, P〈0.05）. It was concluded that plRES-Sj97-Sj 14-Sj26 vaccine may induce stronger immune response in BALB/c mice.