B群腦膜炎球菌外膜蛋白復(fù)合物免疫原性初探
發(fā)布時間：2018-08-19 21:17
【摘要】： B群腦膜炎奈瑟氏球菌是引起流行性腦脊髓膜炎的主要病原菌,其血清型別眾多,型間缺乏交叉免疫保護。B群腦膜炎球菌疫苗的研發(fā)多集中在外膜蛋白(OMPs)及外膜囊(OMV)。自然狀態(tài)的OMV由完整的外膜組成,包括外膜蛋白(OMP)和脂寡糖(LOS)。這些成分很容易通過去垢劑溶解直接從菌體上獲得。 腦膜炎奈瑟氏菌體表面的蛋白質(zhì),依據(jù)分子量可分成5類,其Ⅰ類蛋白是陽離子選擇性孔蛋白,能夠誘導(dǎo)殺菌抗體的產(chǎn)生;Ⅱ類、Ⅲ類蛋白是陰離子選擇性孔蛋白,在一種菌上不能同時表達,抗Ⅱ/Ⅲ類蛋白的抗體只是偶爾具有保護和殺菌活性;Ⅳ類蛋白的功能尚不清楚,可能參與了孔道的形成,抗Ⅳ類蛋白抗體能夠抑制特異的抗Ⅰ類及Ⅱ/Ⅲ類蛋白抗體的殺菌活性,從而大大降低細菌的免疫原性,因此Ⅳ類蛋白不應(yīng)該包含在疫苗成分中,在分離的OMP中盡量減少這種蛋白很重要:V類蛋白在細菌粘附和入侵宿主細胞上起到很大的作用,也是正在被研究的疫苗候選物。 本實驗挑選了3株具有代表性的菌株,使用去垢劑提取了它們的外膜蛋白(OMPs),SDS-PAGE分析發(fā)現(xiàn):3株菌的OMPs帶譜不完全相同,但都含有Ⅰ、Ⅱ/Ⅲ、Ⅳ和V類外膜蛋白。以29356菌株蛋白復(fù)合物的不同劑量和針次免疫NIH小鼠,間接ELISA法檢測小鼠血清IgG抗體滴度,結(jié)果顯示:不同劑量免疫均能誘導(dǎo)小鼠產(chǎn)生IgG抗體,其效價可達1:128000,說明其免疫原性良好。經(jīng)實驗比較選擇最佳免疫方案免疫小鼠,然后用3株腦膜炎球菌菌株進行腹腔攻毒,觀察主動免疫和交叉免疫保護效果,結(jié)果顯示:以10個LD50的活菌攻擊免疫小鼠后,29356菌株組存活率為100%,51169菌株組存活率是0%,29004菌株組存活率是0%,而陰性對照組存活率為0%,表明以該蛋白復(fù)合物免疫小鼠后對同血清型菌的攻毒有很好保護力,但是,對B群的其他菌株缺乏交叉保護。
[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.
【學(xué)位授予單位】：北京生物制品研究所
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R392
本文編號：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.
【學(xué)位授予單位】：北京生物制品研究所
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R392
【相似文獻】
相關(guān)碩士學(xué)位論文 前2條
1 時宇;B群腦膜炎球菌外膜蛋白復(fù)合物免疫原性初探[D];北京生物制品研究所;2009年
2 馬印圖;捕獲包被ELISA方法檢測血小板抗體的可行性研究[D];中國人民解放軍軍事醫(yī)學(xué)科學(xué)院;2006年
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