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首页> 外文期刊>Vaccine >Induction of immune responses by purified outer membrane proteinTI Induction of immune responses by purified outer membrane protein complexes from Neisseria meningitidis
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Induction of immune responses by purified outer membrane proteinTI Induction of immune responses by purified outer membrane protein complexes from Neisseria meningitidis

机译：纯化的外膜蛋白TI诱导免疫应答纯化的脑膜炎奈瑟氏球菌的纯化外膜蛋白复合物诱导免疫应答

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摘要

A broad-spectrum vaccine against disease caused by serogroup B of Neisseria meningitidis is still a challenge due to antigenic variability. In the present study outer membrane protein complexes and their components were analysed using non-denaturing 2D electrophoresis and identified using LC/MS-MS and MALDI-TOF. Outer membrane protein complexes were purified from both the wild-type strain H44/76 and their knock-out mutants lacking PorA, PorB, RmpM or FetA. The immune responses elicited by the whole outer membrane vesicles (OMV) and the purified complexes were analysed for bactericidal activity, antibody surface binding, antibody-mediated C3b/iC3b deposition, membrane attack complex (MAC) deposition and induction of opsonophagocytosis, both on the homologous and several heterologous strains. The main antigenic complexes found were homomeric, formed by the 60 kDa chaperonin (MSP63) or PorB, or heteromeric, formed by different combinations of PorA, PorB and/or RmpM. The lack of some of these proteins in the OMVs from the knock-out mutants did not affect significantly the immune responses analysed except MAC, which was significantly reduced in the anti-PorA and anti-PorB sera, and bactericidal activity, which was absent in the anti-PorA serum. The sera against purified native complexes showed variable activities against the homologous strain, with greatest responses observed for anti-chaperonin and anti-PorA/PorB/RmpM sera. When tested against heterologous strains, the only anti-complex serum showing consistent responses was that against the 60 kDa chaperonin. The comparison of the responses elicited by the different sera suggests an important role of conformational epitopes, present only in native complexes, in the induction of more effective responses against N. meningitidis
机译：由于抗原变异性，针对由脑膜炎奈瑟氏球菌B血清群引起的疾病的广谱疫苗仍然是一个挑战。在本研究中，使用非变性2D电泳分析了外膜蛋白复合物及其成分，并使用LC / MS-MS和MALDI-TOF进行了鉴定。从野生型菌株H44 / 76及其缺少PorA，PorB，RmpM或FetA的敲除突变体中纯化外膜蛋白复合物。分析了整个外膜囊泡（OMV）和纯化的复合物引起的免疫反应的杀菌活性，抗体表面结合，抗体介导的C3b / iC3b沉积，膜攻击复合物（MAC）沉积和调理吞噬作用，两者均在同源和几种异源菌株。发现的主要抗原复合物是由60 kDa伴侣蛋白（MSP63）或PorB形成的同聚体，或由PorA，PorB和/或RmpM的不同组合形成的异聚体。剔除突变体的OMV中缺少某些蛋白质不会显着影响所分析的免疫反应，但MAC的抗-PorA和抗-PorB血清显着降低，而杀菌活性却不存在。抗PorA血清。针对纯化的天然复合物的血清显示出针对同源菌株的可变活性，对于抗伴侣蛋白和抗PorA / PorB / RmpM血清观察到最大的应答。当针对异源菌株进行测试时，唯一显示出一致反应的抗复合血清是针对60 kDa伴侣蛋白的。不同血清引起的应答的比较表明，仅在天然复合物中存在的构象表位在诱导针对脑膜炎奈瑟氏球菌的更有效应答中具有重要作用。

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