论文部分内容阅读
主动获得性免疫已经帮助人类彻底战胜了天花,并且大幅度降低了脊髓灰质炎和麻疹等多种疾病的发病率和死亡率,但是现行的亚单位疫苗至少有两点不足之处:(1)只能将病毒的一部分提供给免疫系统,免疫效果仍然不够理想,病毒感染(例如HIV等)仍然是造成人类残疾和死亡的主要因素之一.(2)亚单位疫苗采用蛋白质抗原,或者利用病毒做载体,制备复杂,成本高.因此人类面临开发新一代高效低成本疫苗的挑战.基因免疫(Genetic immunization)是基于裸露DNA转基因成功这一最新发现而发展起来的新的免疫技术,相应的疫苗称之为基因疫苗(Genetic vaccine).同亚单位疫苗相比,基因疫苗具有安全性高、免疫效果好、免疫作用持久和制备方便等优点,被誉为疫苗史上的革命.我们选用乙型肝炎病毒表面抗原(HBsAg)开展基因免疫研究,通过基因枪介导将HBsAg表达质粒转入小鼠耳部,组织切片观察基因转入位点,测小鼠血清中特异抗体确定基因免疫效果.1 材料与方法1.1 实验材料带有人β-肌动蛋白启动子的表达载体由美国西南医学中心Johnston教授惠赠,HBsAg表达质粒由本研究室自由构建(图1).抗-HBs抗体检测采用上海生物制品所生产的试剂盒,C57BL/6和昆明小鼠购自中国科学院动物研究所,羊抗鼠IgG-HRP,由北京生物制品所生产.基因枪由清华大学生?
Actively acquired immunity has helped to completely defeat smallpox and greatly reduce the morbidity and mortality of many diseases such as poliomyelitis and measles. However, the current subunit vaccines have at least two deficiencies: (1) Only a part of the virus can be provided to the immune system, the immune effect is still not ideal, and virus infection (such as HIV, etc.) remains one of the major causes of human disability and death. (2) Subunit vaccines use protein antigens, Therefore, human beings are faced with the challenge of developing a new generation of high-efficiency and low-cost vaccines.Genetic immunization is a new immunological technology developed based on the latest discovery of the success of naked DNA transgene, and the corresponding vaccine Called genetic vaccine (Genetic vaccine) .Compared with the subunit vaccine, gene vaccine has the advantages of high safety, good immune effect, long lasting immunization and easy preparation, etc. It is regarded as the revolution in the history of vaccine.We choose hepatitis B Virus surface antigen (HBsAg) to carry out genetic immunization studies, HBsAg expression plasmid mediated by gene gun transferred to the ear, tissue cutting Observe the gene transfer site and measure the specific antibody in serum to confirm the effect of gene immunization.Materials and Methods1.1 Experimental Materials The expression vector with humanβ-actin promoter was kindly donated by Professor Johnston from Southwestern Medical Center of the United States. The HBsAg expression plasmid (Figure 1) Anti-HBs antibody detection using Shanghai biological products produced kits, C57BL / 6 and Kunming mice purchased from the Institute of Zoology, Chinese goat anti-mouse IgG-HRP by the Beijing Institute of Biology Products produced by gene gun by Tsinghua University students?