弓形虫SAG1、GRA2单基因疫苗及SAG1-GRA2复合基因疫苗的免疫效果观察  

Observation on the immune responses of the monvalent SAG1,GRA2 and compound SAG1-GRA2 DNA vaccine against Toxoplasma gondii

作　　者：李首庆[1] 古钦民[1] 杨广民[1] 马寅芙[1] 

机构地区：[1]吉林省人民医院中心实验室,吉林长春130021

出　　处：《中国实验诊断学》2008年第10期1219-1222,共4页Chinese Journal of Laboratory Diagnosis

基　　金：山东省自然科学基金项目(No.Y2005C20);山东省医药卫生科技发展项目(No.2005HZ028)

摘　　要：目的检测SAG1和GRA2基因编码的单基因及复合基因疫苗诱导小鼠的免疫应答,评价其抗弓形虫感染的保护性免疫效果。方法大量制备pcDNA3.1-SAG1、pcDNA3.1-GRA2、pcDNA3.1-SAG1-GRA2真核表达重组质粒,肌内注射BALB/c小鼠,分别于2周、4周后加强免疫一次。另设立pcDNA3.1及PBS对照组。于3次免疫后4周作免疫指标测定(包括ELISA法测定小鼠血清IgG抗体及细胞因子IFN-γ、IL-4,MTT法测定小鼠淋巴细胞增殖能力及NK细胞杀伤活性),并观察弓形虫速殖子攻击感染后小鼠的生存时间。结果SAG1-GRA2组小鼠血清IgG抗体、IFN-γ、淋巴细胞增殖能力及NK细胞杀伤活性均高于SAG1及GRA2组(P<0.05);各组均未检测到IL-4;复合基因组小鼠生存时间较单基因疫苗组延长(P<0.01)。结论弓形虫SAG1-GRA2复合基因疫苗较SAG1、GRA2单基因疫苗具有更好的免疫保护性。Objective To examine the immune response by monvalent SAG1, GRA2 and compound SAG1-GRA2 DAN vaccine and assess the protective effect against Toxoplasmosis.Methods Abundant plasmids were extracted and BALB/c mice were intramus-cularly-immunized with the DNA vaccines, two booster injection were carried out at 2-week intervals .The empty vector peDNA3.1 and PBS buffer were as negative controls. Four weeks after the third inmmnization, the following tests were conducted： IgG, IFN-Tand IL-4 were determined by means of ELISA, the proliferative activity of T lyrnphcytes and the killing activity of NK cells were detected by means of MTr assay. Then, analyse the survival time of mice infected by Toxoplasma gondii. Results The value of IgG, IFN-γ and the activity of T lymph cells and NK cells were all higher in the compound group than the both monogenic groups, the value of IL-4 was not sighnificant, the living time of mice in the compound group was prolonged obviously compared with other goups. Conclusion SAG1-GRA2 DNA vaccine elicits better immunprotecfivity than SAG1, GRA2 DNA vaccine in mice.

关 键 词：弓形虫 SAG1 GRA2 复合基因 疫苗 

分 类 号：R392[医药卫生—免疫学]