B群脑膜炎球菌外膜蛋白复合物免疫原性初探
发布时间：2018-08-19 21:17
【摘要】： B群脑膜炎奈瑟氏球菌是引起流行性脑脊髓膜炎的主要病原菌,其血清型别众多,型间缺乏交叉免疫保护。B群脑膜炎球菌疫苗的研发多集中在外膜蛋白(OMPs)及外膜囊(OMV)。自然状态的OMV由完整的外膜组成,包括外膜蛋白(OMP)和脂寡糖(LOS)。这些成分很容易通过去垢剂溶解直接从菌体上获得。 脑膜炎奈瑟氏菌体表面的蛋白质,依据分子量可分成5类,其Ⅰ类蛋白是阳离子选择性孔蛋白,能够诱导杀菌抗体的产生;Ⅱ类、Ⅲ类蛋白是阴离子选择性孔蛋白,在一种菌上不能同时表达,抗Ⅱ/Ⅲ类蛋白的抗体只是偶尔具有保护和杀菌活性;Ⅳ类蛋白的功能尚不清楚,可能参与了孔道的形成,抗Ⅳ类蛋白抗体能够抑制特异的抗Ⅰ类及Ⅱ/Ⅲ类蛋白抗体的杀菌活性,从而大大降低细菌的免疫原性,因此Ⅳ类蛋白不应该包含在疫苗成分中,在分离的OMP中尽量减少这种蛋白很重要:V类蛋白在细菌粘附和入侵宿主细胞上起到很大的作用,也是正在被研究的疫苗候选物。 本实验挑选了3株具有代表性的菌株,使用去垢剂提取了它们的外膜蛋白(OMPs),SDS-PAGE分析发现:3株菌的OMPs带谱不完全相同,但都含有Ⅰ、Ⅱ/Ⅲ、Ⅳ和V类外膜蛋白。以29356菌株蛋白复合物的不同剂量和针次免疫NIH小鼠,间接ELISA法检测小鼠血清IgG抗体滴度,结果显示:不同剂量免疫均能诱导小鼠产生IgG抗体,其效价可达1:128000,说明其免疫原性良好。经实验比较选择最佳免疫方案免疫小鼠,然后用3株脑膜炎球菌菌株进行腹腔攻毒,观察主动免疫和交叉免疫保护效果,结果显示:以10个LD50的活菌攻击免疫小鼠后,29356菌株组存活率为100%,51169菌株组存活率是0%,29004菌株组存活率是0%,而阴性对照组存活率为0%,表明以该蛋白复合物免疫小鼠后对同血清型菌的攻毒有很好保护力,但是,对B群的其他菌株缺乏交叉保护。
[Abstract]:Neisseria meningitidis group B is the main pathogen causing epidemic cerebrospinal meningitis. It has many serotypes and lacks cross-immune protection. The development of group B meningococcal vaccine mainly focuses on outer membrane protein (OMPs) and outer capsule (OMV). Natural OMV consists of intact outer membrane, including outer membrane protein (OMP) and Lipooligosaccharide (LOS). These components can be easily obtained from bacteria by dissolution of detergent.
Proteins on the surface of Neisseria meningitidis can be classified into five categories according to their molecular weight. Cationic selective porins are class I proteins that can induce the production of bactericidal antibodies; class II and class III proteins are anionic selective porins that can not be expressed simultaneously in one strain, and antibodies against class II/III proteins are only occasionally protective and bactericidal. The function of class IV protein is not clear and may be involved in pore formation. Anti-class IV protein antibodies can inhibit the bactericidal activity of specific antibodies against class I and class II/III proteins, thus greatly reducing the immunogenicity of bacteria. Therefore, class IV proteins should not be included in vaccine components and should be minimized in isolated OMP. Important: V proteins play an important role in bacterial adhesion and invasion of host cells, and are also being studied as vaccine candidates.
Three representative strains were selected and their outer membrane proteins (OMPs) were extracted by detergent. SDS-PAGE analysis showed that the OMPs bands of the three strains were not identical, but all contained outer membrane proteins of class I, II/III, IV and V. The titer of serum IgG antibody showed that different doses of immunization could induce the production of IgG antibody in mice. The titer of IgG antibody reached 1:128000, indicating that the immunogenicity was good. The results showed that the survival rates of 29356, 51169, 29004 and negative control groups were 100%, 0% and 0% respectively after inoculating mice with 10 LD50 live bacteria, indicating that the protein complex could protect mice against the same serotype of bacteria, but the other strains of group B were not cross-linked. Protection.
【学位授予单位】：北京生物制品研究所
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392
本文编号：2192852
[Abstract]:Neisseria meningitidis group B is the main pathogen causing epidemic cerebrospinal meningitis. It has many serotypes and lacks cross-immune protection. The development of group B meningococcal vaccine mainly focuses on outer membrane protein (OMPs) and outer capsule (OMV). Natural OMV consists of intact outer membrane, including outer membrane protein (OMP) and Lipooligosaccharide (LOS). These components can be easily obtained from bacteria by dissolution of detergent.
Proteins on the surface of Neisseria meningitidis can be classified into five categories according to their molecular weight. Cationic selective porins are class I proteins that can induce the production of bactericidal antibodies; class II and class III proteins are anionic selective porins that can not be expressed simultaneously in one strain, and antibodies against class II/III proteins are only occasionally protective and bactericidal. The function of class IV protein is not clear and may be involved in pore formation. Anti-class IV protein antibodies can inhibit the bactericidal activity of specific antibodies against class I and class II/III proteins, thus greatly reducing the immunogenicity of bacteria. Therefore, class IV proteins should not be included in vaccine components and should be minimized in isolated OMP. Important: V proteins play an important role in bacterial adhesion and invasion of host cells, and are also being studied as vaccine candidates.
Three representative strains were selected and their outer membrane proteins (OMPs) were extracted by detergent. SDS-PAGE analysis showed that the OMPs bands of the three strains were not identical, but all contained outer membrane proteins of class I, II/III, IV and V. The titer of serum IgG antibody showed that different doses of immunization could induce the production of IgG antibody in mice. The titer of IgG antibody reached 1:128000, indicating that the immunogenicity was good. The results showed that the survival rates of 29356, 51169, 29004 and negative control groups were 100%, 0% and 0% respectively after inoculating mice with 10 LD50 live bacteria, indicating that the protein complex could protect mice against the same serotype of bacteria, but the other strains of group B were not cross-linked. Protection.
【学位授予单位】：北京生物制品研究所
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392
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相关硕士学位论文 前2条
1 时宇;B群脑膜炎球菌外膜蛋白复合物免疫原性初探[D];北京生物制品研究所;2009年
2 马印图;捕获包被ELISA方法检测血小板抗体的可行性研究[D];中国人民解放军军事医学科学院;2006年
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