Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom.
PLoS Pathog. 2012;8(10):e1002981. doi: 10.1371/journal.ppat.1002981. Epub 2012 Oct 25.
Neisseria meningitis remains a leading cause of sepsis and meningitis, and vaccines are required to prevent infections by this important human pathogen. Factor H binding protein (fHbp) is a key antigen that elicits protective immunity against the meningococcus and recruits the host complement regulator, fH. As the high affinity interaction between fHbp and fH could impair immune responses, we sought to identify non-functional fHbps that could act as effective immunogens. This was achieved by alanine substitution of fHbps from all three variant groups (V1, V2 and V3 fHbp) of the protein; while some residues affected fH binding in each variant group, the distribution of key amino underlying the interaction with fH differed between the V1, V2 and V3 proteins. The atomic structure of V3 fHbp in complex with fH and of the C-terminal barrel of V2 fHbp provide explanations to the differences in the precise nature of their interactions with fH, and the instability of the V2 protein. To develop transgenic models to assess the efficacy of non-functional fHbps, we determined the structural basis of the low level of interaction between fHbp and murine fH; in addition to changes in amino acids in the fHbp binding site, murine fH has a distinct conformation compared with the human protein that would sterically inhibit binding to fHbp. Non-functional V1 fHbps were further characterised by binding and structural studies, and shown in non-transgenic and transgenic mice (expressing chimeric fH that binds fHbp and precisely regulates complement system) to retain their immunogenicity. Our findings provide a catalogue of non-functional fHbps from all variant groups that can be included in new generation meningococcal vaccines, and establish proof-in-principle for clinical studies to compare their efficacy with wild-type fHbps.
脑膜炎奈瑟菌仍然是脓毒症和脑膜炎的主要原因,需要疫苗来预防这种重要的人类病原体感染。因子 H 结合蛋白 (fHbp) 是一种关键抗原,可引发针对脑膜炎奈瑟球菌的保护性免疫,并募集宿主补体调节剂 fH。由于 fHbp 与 fH 之间的高亲和力相互作用可能会损害免疫反应,因此我们试图鉴定可作为有效免疫原的非功能 fHbp。这是通过对该蛋白的所有三个变体组(V1、V2 和 V3 fHbp)中的 fHbp 进行丙氨酸取代来实现的;虽然某些残基在每个变体组中都影响 fH 结合,但与 fH 相互作用的关键氨基酸的分布在 V1、V2 和 V3 蛋白之间存在差异。V3 fHbp 与 fH 复合物的原子结构和 V2 fHbp 的 C 末端桶的原子结构为它们与 fH 相互作用的精确性质以及 V2 蛋白的不稳定性的差异提供了解释。为了开发转基因模型来评估非功能 fHbp 的功效,我们确定了 fHbp 与鼠 fH 之间低水平相互作用的结构基础;除了 fHbp 结合位点中的氨基酸变化外,鼠 fH 与人类蛋白的构象明显不同,这会在空间上抑制与 fHbp 的结合。通过结合和结构研究进一步表征非功能 V1 fHbp,并在非转基因和转基因小鼠(表达与 fHbp 结合并精确调节补体系统的嵌合 fH)中显示,它们保留了免疫原性。我们的研究结果提供了所有变体组中非功能 fHbp 的目录,这些 fHbp 可包含在新一代脑膜炎球菌疫苗中,并为临床研究提供了原理证明,以比较它们与野生型 fHbp 的功效。