CONSERVED NEISSERIA PROTEINS AS VACCINE CANDIDATES

描述：(改编自申请者摘要) 这项研究是为了增加我们对保存膜的使用的理解 脑膜炎奈瑟氏菌预防疫苗中的蛋白质成分 B群(MenB)病。乙型脑膜炎是脑膜炎和败血症的主要原因。 尽管血清杀菌抗体提供了保护，但到目前为止，传统的 开发疫苗的方法在很大程度上并不成功。 以多糖为基础的MenB疫苗有可能引发针对宿主的自身抗体 聚唾液酸，而大多数非囊性抗原诱导 基础广泛的免疫受到抗原多样性的限制。我们建议 研究最近发现的三种保守的疫苗潜力 NeisSeries膜蛋白，命名为NeisSeries表面蛋白(NSP)A， B和C。作为备用候选，NSPD和NSPE也可用。NSPA是 用一种单抗发现，而其他四种蛋白质代表 通过对基因组数据的分析发现了新的候选疫苗。全 五种蛋白在致病奈瑟氏菌中高度保守，具有表位 在细菌的表面上，抗体可以接触到，并诱导 补体介导的杀菌抗体在小鼠或兔体内。因此，每一个 这些蛋白质作为人类免疫缺陷病毒的候选抗原值得进一步研究 包括在MenB疫苗中。在目标1中，我们将研究其免疫原性。 每一种重组蛋白在小鼠和豚鼠中的表达。该不该 重组分子不能诱导高滴度的抗体 在对抗细菌的功能上，我们将尝试重建 使用洗涤剂或脂质体的构象表位，并探索 使用适合人类使用的新型佐剂。在目标2中，我们将准备 与NS蛋白上的表位发生反应的单抗 在诱导保护性抗体方面很重要。这些单抗将用于 表位映射，以及抗体功能活性的研究。在目标3中，我们 也将使用3Iabs来调查是否存在菌株差异 不同NS蛋白的表面可及性和表达，以及 将发现的任何差异与相应的DNA序列编码 蛋白质，或各自基因的转录活性。我们也会 研究不同NS蛋白的表面可及性是否有所不同 当在体外繁殖时，在NeisSeries菌株内，或在幼鼠体内。在AIM 4，我们将测试一种包含多个NS的疫苗的假设 与疫苗相比，蛋白质将产生更广泛的对MenB病毒的保护性免疫 来自一种单一蛋白质。这些结果与评估 在MenB中包含或排除这些新蛋白的可能性 疫苗。该数据还可以验证用于鉴定的基因组方法 疫苗开发的新抗原性靶点。

DESCRIPTION: (Adapted from Applicant's Abstract) The long-term objective of this study is to increase our understanding of the use of conserved membrane proteins as components of a vaccine for prevention of Neisseria meningitidis serogroup B (MenB) disease. MenB is a major cause of meningitis and sepsis. Although serum bactericidal antibodies confer protection, to date, conventional approaches to develop a vaccine have been largely unsuccessful. Polysaccharide-based MenB vaccines risk eliciting autoantibodies to host polysialic acid, while the ability of most non-capsular antigens to elicit broad-based immunity is limited by antigenic diversity. We propose to investigate the vaccine potential of three recently discovered conserved Neisserial membrane proteins, designated Neisserial surface proteins (Nsp) A, B, and C. As backup candidates, NspD and NspE are also available. NspA was discovered with a monoclonal antibody, while the other four proteins represent new vaccine candidates that were discovered from analysis of genomic data. All five proteins are highly conserved across pathogenic Neisseria, have epitopes on the surface of the bacteria that are accessible to antibody, and elicit complement-mediated bactericidal antibodies in mice or rabbits. Thus, each of these proteins deserves further investigation as candidate antigens for inclusion in a MenB vaccine. In Aim 1, we will investigate the immunogenicity of each of the recombinant proteins in mice and guinea pigs. Should the recombinant molecules fail to elicit high titers of antibodies that are functionally active against the bacteria, we will attempt to reconstitute conformational epitopes with the use of detergents or liposomes, and explore the use of novel adjuvants suitable for human use. In Aim 2, we will prepare monoclonal antibodies (Mabs) that react with epitopes on the Ns proteins that are important in eliciting protective antibodies. These Mabs will be used for epitope mapping, and for studies of antibody functional activity. In Aim 3, we also will use the 3Iabs to investigate whether there are strain differences in surface accessibility and expression of the different NS proteins, and correlate any differences found with the respective DNA sequences encoding the proteins, or transcriptional activity of the respective genes. We also will investigate whether surface accessibility of the different Ns proteins varies within a Neisserial strain when propagated in vitro, or in infant rats. In Aim 4, we will test the hypothesis that a vaccine containing more than one Ns protein will elicit broader protective immunity to MenB than a vaccine made from a single protein. These results are directly relevant to evaluating the potential for inclusion or exclusion of each of these novel proteins in a MenB vaccine. The data also may validate the genomic approach for identification of new antigenic targets for vaccine development.