猪流产嗜性衣原体omp-1基因的重组质粒与重组蛋白免疫组合效果  

Evaluation of immune response of different combinations of swine Chlamydophila abortus omp-1 and its recombinant protein

作　　者：杨琪[1] 李建平[2] 杨建民[1] 李军[1] 杨利[1] 何诚[1] 

机构地区：[1]中国农业大学动物医学院,北京100094 [2]中央民族大学体育系,北京100086

出　　处：《中国农业大学学报》2007年第3期9-13,共5页Journal of China Agricultural University

基　　金：国家自然科学基金资助项目(30370070);北京市自然科学基金资助项目(6052014)

摘　　要：为增强猪流产嗜性衣原体omp-1基因的免疫措施进而筛选出高效新型分子疫苗,本实验将猪流产嗜性衣原体omp-1基因克隆入pcDNA3.1(+)载体构建DNA疫苗,初次和二次免疫分别以核酸/重组蛋白(DNA/r-MOMP)、重组蛋白/核酸(r-MOMP/DNA)、重组蛋白+核酸/重组蛋白+核酸(r-MOMP+DNA/r-MOMP+DNA)、核酸+核酸(DNA/DNA)和重组蛋白+重组蛋白(r-MOMP/r-MOMP)5种组合接种BALB/c小鼠,同时以载体和弱毒为对照。二次免疫后14 d以ELISA和T淋巴细胞增殖实验检测特异性体液免疫和细胞免疫应答水平。结果显示核酸与重组蛋白同时免疫可以诱导产生较高的体液免疫和细胞免疫水平,而DNA/r-MOMP组合的细胞免疫和体液免疫水平高于r-MOMP/DNA组合,单独免疫DNA组或r-MOMP都不能获得好的细胞和体液免疫。The omp-1 DNA is known to be a strong immunogen and codes the 40 ku major outer-membrane protein （MQMP）, an immunodominant antigen of the Chlarnydiaceae surface membrane. A reliable and efficient molecular vaccine against swine Chiamydophila abortus Cp. abortus can be developed if a new approach for increasing immune response can be found, In the current study, the omp-1 of CP12 strain of Cp. abortus was amplified by PCR and cloned into pcDNA3.1（ ＋ ）to construct recombinant eukaryotic expression vector. BALB/c mice were randomly divided into five groups and inoculated twice weekly with different combinations such as DNA/r-MOMP, r-MOMF;/DNA, DNA-r- MOMP, DAN/DNA and r-MOMP/r-MOMP. Two weeks post inoculation, the antibodies against MOMP were assessed by ELISA, and proliferation of T lymphocytes was monitored by Lymphocyte Transformation Test. Results showed that sig- nificantly higher immunoreactions could be induced in mice inoculated simultaneously with DNA and r-MOMP. Mice group primed with DNA and then boosted with r-MOMP exhibited higher level antibody and lymphocytes stimulation in- dex as than that inoculated with r-MOMP, then with DNA. Reliable and efficient immunoreactions cannot be achieved ei- ther by inoculation with naked DNA or with recombinant MOMP. Nevertheless, a better immune response can be obtained if mice are administered with both DNA and its corresponding recombinant protein simultaneously.

关 键 词：猪流产嗜性衣原体 omp-1基因 重组质粒 重组蛋白 免疫 小鼠 

分 类 号：S852.76[农业科学—基础兽医学]