炭疽芽孢苗与吸附PA苗诱导小鼠体液免疫应答初步观察

发布时间：2018-05-12 00:31

本文选题：吸附PA苗 + A16R　；参考：《中国人民解放军军事医学科学院》2007年硕士论文

【摘要】： 炭疽芽孢杆菌是人畜共患炭疽病的重要病原体，是公认的重要生物战剂之一，目前人用疫苗有两种：即炭疽减毒活芽孢苗和炭疽弱毒株无菌培养滤液佐剂吸附苗，自研制成功以来，已在人群中长期使用，为了深入了解炭疽疫苗的免疫应答机制，为疫苗的开发及评价提供理论基础，本研究采用炭疽芽孢杆菌两种疫苗——A16R芽孢苗与吸附PA苗，，分别从三个层面上——脾、骨髓和外周血，初步观察炭疽两种疫苗的免疫应答机制。本研究共分为三部分：1、炭疽两种疫苗的制备及免疫效果测定，分两部分(1)两种炭疽疫苗的制备和鉴定；(2)A16R芽孢苗及吸附PA苗免疫Balb／C小鼠血清抗体效价动态观察及抗体亚型分析；2、A16R芽孢苗及吸附PA苗诱导Balb／C小鼠特异性ASCs的动态观察(1)首先建立了ELISPOT检测炭疽疫苗免疫Balb／C小鼠后抗原特异性抗体分泌细胞的实验方法；(2)从三个层面上，即骨髓、脾和外周血，观察了两种疫苗免疫Balb／C小鼠后特异性ASCs的动态变化及差异性，并结合外周血抗体效价的消长分析不同器官／部位ASCs应答与外周血抗体效价消长之间的相关性；3、炭疽A16R芽孢苗与吸附PA苗诱导Balb／C小鼠脾、骨髓及外周血中特异性Th2细胞应答的动态变化，并结合外周血抗体效价的消长分析不同器官／部位Th2应答的产生与维持对稳定血中抗芽孢／PA抗体的辅助作用。结论：(1)通过对炭疽两种疫苗免疫后Balb／C小鼠外周血抗体效价的动态观察，发现随着两种疫苗的加强免疫，小鼠血清中均可检测到高效价的抗体水平，并维持较长时间；吸附PA苗免疫组抗体亚型分析以IgG1和IgG2b为主，IgG2a水平较低，与对照组差异显著；A16R芽孢苗免疫组抗体亚型分析虽仍以IgG1和IgG2b为主，亦可检测到较高水平的IgG2a；(2)两种疫苗免疫后Balb／C小鼠脾、骨髓和血中均可检测到ASCs，但ASCs出现时间及强度不同，脾中最先出现，首先达峰值，骨髓中峰值晚于脾，但骨髓中应答维持时间较长，与外周血抗体效价的维持有一定的相关性；两种疫苗免疫后Balb／C小鼠脾、骨髓及外周血中ASCs的动态变化有一定差异；(3)两种疫苗免疫后Balb／C小鼠脾、骨髓和外周血中均可检测到抗原特异性Th2细胞应答，但Th2应答出现时间及强度不同，骨髓中最先出现，首先达峰值，且维持时间长，外周血中可检测到较强的Th2应答，且随着脾、骨髓及外周血的Th2应答的升高与维持，外周血抗体滴度不断升高，且于末次加强免疫后60天或30天仍维持在较高水平。疫苗免疫应答是一个复杂的多层次的过程，受复杂的网络调控，因此对于疫苗的评价应从多方面、多角度入手，全面、深入、系统的研究。
[Abstract]:Bacillus anthracis is an important pathogen of zoonotic anthracnose and one of the most important biological warfare agents. At present, there are two kinds of human vaccines: anthrax attenuated live spores vaccine and anthrax attenuated strain sterile culture filtrate adjuvant adsorbent vaccine. Since the successful development of anthrax vaccine, it has been used for a long time. In order to understand the immune response mechanism of anthrax vaccine and provide the theoretical basis for vaccine development and evaluation, In this study, two kinds of Bacillus anthracis vaccine, Bacillus anthracis A16R vaccine and adsorbed PA vaccine, were used to observe the immune response mechanism of the two vaccines from three levels: spleen, bone marrow and peripheral blood, respectively. This study was divided into three parts: the preparation of two anthrax vaccines and the determination of their immune effects. Preparation and Identification of two anthrax Vaccines: dynamic observation of Serum Antibody titer and Antibody subtype Analysis of Balb/C mice immunized with A16R Bacillus Vaccine and Adsorbed PA Vaccine; dynamic observation of specific ASCs in Balb/C mice induced by A16R Bacillus Vaccine and Adsorbed PA Vaccine In this paper, we first established a ELISPOT method for the detection of antigen-specific antibody secreting cells in Balb/C mice immunized with anthrax vaccine. Bone marrow, spleen and peripheral blood were used to observe the dynamic changes and differences of specific ASCs in Balb/C mice immunized with two vaccines. The correlation between ASCs response and peripheral blood antibody titer in different organs / parts was analyzed. The spleen of Balb/C mice was induced by anthrax A16R spores and PA adsorbed vaccine. The dynamic changes of specific Th2 cell responses in bone marrow and peripheral blood, and the auxiliary effect of Th2 response in different organs / parts on the anti-spores / PA antibody in stable blood were analyzed in combination with the increase and decline of antibody titers in peripheral blood. Conclusion by observing the antibody titers in peripheral blood of Balb/C mice immunized with two anthrax vaccines, it was found that the antibody level of high titer could be detected in serum of mice with the enhanced immunization of the two vaccines and maintained for a long time. The level of IgG2a in the immunized group was lower than that in the control group, but IgG1 and IgG2b were still the main subtypes of the antibody subtype in the immunized group of the adsorbed PA vaccine, and the difference was significant compared with that of the control group, and the antibody subtypes of the immunized group were higher than that of the control group. ASCs were also detected in spleen, bone marrow and blood of Balb/C mice immunized with two kinds of vaccines, but the onset time and intensity of ASCs were different, the first appeared in spleen, and the peak value in bone marrow was later than that in spleen. However, the duration of response in bone marrow was longer, which was related to the maintenance of antibody titer in peripheral blood, and the dynamic changes of ASCs in spleen, bone marrow and peripheral blood of Balb/C mice after immunization with the two vaccines were different. Antigen-specific Th2 cell responses were detected in both bone marrow and peripheral blood, but the onset time and intensity of Th2 response were different, the first peak appeared in bone marrow, and the maintenance time was long. A strong Th2 response could be detected in peripheral blood. With the increase and maintenance of Th2 response in spleen, bone marrow and peripheral blood, the antibody titer of peripheral blood increased continuously, and remained at a high level 60 or 30 days after the last booster immunization. Vaccine immune response is a complex and multi-level process, which is regulated by complex network. Therefore, the evaluation of vaccine should be carried out from many aspects, angles, comprehensive, in-depth and systematic research.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R392

【共引文献】