| 摘要: |
| 本研究将实验室重组构建并已验证具有良好的反应原性与免疫原性的两种核酸疫苗(去除428-446序列的pcDNA3.1-ADV-428和同时去除428-446和487-501序列的pcDNA3.1-ADV-428-487)经肌肉注射接种到水貂体内进行免疫。免疫完成后对水貂进行攻毒,然后每隔两周对水貂进行指尖采血。应用流式细胞术检测全血中的CD8+淋巴细胞亚群；应用间接ELISA法检测血清中的抗体水平；应用血清蛋白电泳检测血清中γ球蛋白占总蛋白的百分比以及应用聚乙二醇沉淀比浊法检测水貂血清中抗原抗体复合物的含量。通过上述实验判定pcDNA3.1-ADV-428和 pcDNA3.1-ADV-428-487对水貂阿留申病的防治效果。最后实验结果表明pcDNA3.1-ADV-428和 pcDNA3.1-ADV-428-487的各项数据都要优于对照组,且pcDNA3.1-ADV-428-487的保护效果要优于pcDNA3.1-ADV-428,两种核酸疫苗都展现了良好的疫苗潜质。 |
| 关键词: 水貂阿留申病 核酸疫苗 免疫 抗原抗体复合物 γ球蛋白 淋巴细胞 |
| DOI： |
| 投稿时间：2017-03-07修订日期：2017-11-23 |
| 基金项目:国家自然基金(NO.31272565) |
|
| Preliminary Immune effect evaluation of two ADV recombinant DNA vaccines |
|
| (Jilin Agricultural University) |
| Abstract: |
| In this study, we constructed and validated the laboratory two recombinant nucleic acid vaccine has good reactionogenicity and immunogenicity by After intramuscular inoculation to mink body immune (removal of 428-446 sequence of pcDNA3.1-ADV-428 and simultaneous removal of 428-446 and 487-501 sequences of pcDNA3.1-ADV-428-487). Mink were challenged experiment when after the completion of immunization, then every two weeks for blood collection of mink fingertips. the whole blood CD8+ lymphocyte subsets detection by flow cytometry; Detection of antibody levels in serum by indirect ELISA; Serum protein electrophoresis was used to detect the γ globulin percentage of total protein in serum and the application of polyethylene glycol precipitation determination of antigen antibody complexes in serum of mink. Through the above experimental determination pcDNA3.1-ADV-428 and pcDNA3.1-ADV-428-487 have effect on the prevention and cure of Aleutian disease of mink. Finally, the experimental results show that the data of pcDNA3.1-ADV-428 and pcDNA3.1-ADV-428-487 were better than those of the control group, the protective effect of pcDNA3.1-ADV-428-487 is better than pcDNA3.1-ADV-428, two nucleic acid vaccines have shown good vaccine potential. |
| Key words: Aleutian disease Nucleic acid vaccine Immunization Antigen antibody complex γ globulin lymphocyte |
