Project-002

该项目的目标是开发一种广泛的抗淋球菌交叉保护疫苗， 淋病是一种常见的性传播疾病。N. 对大多数抗生素具有耐药性的淋病引起了人们对无法治愈的淋病的担忧， 迫切需要开发和实施有效的疫苗来预防这种感染及其 后遗症N.淋病是人类特异性病原体，仅存在于泌尿生殖道或更少 通常，在其人类宿主的上呼吸道。我们的方法是将目标对准表面成分 负责介导从宿主铁结合蛋白、转铁蛋白（Tf）和乳铁蛋白（Lf）获得铁， 由于这两个系统已被证明是必不可少的生存的细菌和他们的能力， 感染在人类男性尿道感染模型中的作用。由于细菌没有这些受体就无法生存， 蛋白质，他们将无法逃脱疫苗覆盖的受体损失，因此可以消除 如果我们成功地开发出一种完全交叉保护的疫苗， 受体蛋白的变体。 本项目的重点是利用结构导向的抗原设计来产生靶向于 表面脂蛋白、乳铁蛋白结合蛋白B（Lbp B）和完整的外膜转运 蛋白质，乳铁蛋白结合蛋白A（LbpA），将共同诱导广泛的交叉保护性免疫反应。 针对这些蛋白质的所有已知变体的反应。我们的新型整合疫苗设计和评估 管道方法将在免疫测定中使用抗原变体文库，在定殖中使用菌株文库 和感染模型，以评估免疫应答的交叉反应和交叉保护特性， 指导抗原组合的选择。我们将确定目标的全局序列多样性 抗原（LbpB和LbpA），然后开发代表性变体LbpB的非结合突变体， 预测共同提供针对所有LbpB变体的广泛交叉保护性免疫应答。我们将 评价LbpB的C-叶中带负电荷区域的免疫学特性，确定LbpB的C-叶中带负电荷区域的免疫学特性。 它们的去除的影响，并确定LbpA表位的最佳组合，可以展示在 LbpB C-叶，其诱导针对LbpA的广泛交叉保护应答。 最后一步将是整合来自一个已建立的NIH拨款的结果，该拨款专注于开发抗原 靶向转铁蛋白结合蛋白B和A（Tbp B和TbpA），并确定以下的最佳组合： 工程化的TbpB和LbpB抗原展示来自TbpA和LbpA的表位，以诱导广泛的交叉保护性免疫应答。 对所有四种抗原的免疫反应。在我们的正常和转基因小鼠中的研究表明， 靶向TbpB或TbpA的抗原可以减少定殖，因此开发疫苗的前景是， 消除表达四种靶抗原中任何一种的细菌是有希望的。

This goal of this project is to develop a broadly cross-protective vaccine against Neisseria gonorrhoeae, the bacteria responsible for the common sexually-transmitted disease, gonorrhea. The emergence of strains of N. gonorrhoeae that are resistant to most antibiotics has raised the specter of untreatable gonorrhea, adding urgency for the development and implementation of effective vaccines for prevention of this infection and its sequelae. N. gonorrhoeae is human-specific pathogen that exclusively resides in genitourinary tract, or less frequently, in the upper respiratory tract, of its human host. Our approach is to target the surface components responsible for mediating iron acquisition from the host iron binding proteins, transferrin (Tf) and lactoferrin (Lf), since these two systems have been shown to be essential for survival of the bacteria and their ability to cause infection in a human male urethral infection model. Since the bacteria cannot survive without these receptor proteins, they will not be able to escape vaccine coverage by loss of the receptors, thus could be eliminated from vaccinated individuals if we are successful in developing a fully cross-protective vaccine against all variants of the receptor proteins. This project is focused on using structure-guided antigen design to generate engineered antigens targeting both the surface lipoprotein, lactoferrin binding protein B (LbpB), and the integral outer membrane transport protein, lactoferrin binding protein A (LbpA), that will collectively induce a broadly cross-protective immune response against all known variants of these proteins. Our novel integrated vaccine design and evaluation pipeline approach will use libraries of antigenic variants in immunoassays and strain libraries in colonization and infection models to evaluate the cross-reactive and cross protective properties of the immune response to guide the selection of combinations of antigens. We will determine the global sequence diversity of the target antigens (LbpB and LbpA) and then develop non-binding mutants of representative variant LbpBs that are predicted to collectively provide a broad cross-protective immune response against all LbpB variants. We will evaluate the immunological properties of the negatively charged regions in the C-lobe of LbpB, determine the impact of their removal and determine the optimum combination of LbpA epitopes that can be displayed on the LbpB C-lobe that induces a broadly cross-protective response against LbpA. The final step will be to integrate the results from an established NIH grant focused on developing antigens targeting transferrin binding protein B and A (TbpB and TbpA) and determine the optimum combination of engineered TbpB and LbpB antigens displaying epitopes from TbpA and LbpA to induce a broadly cross-protective immune response against all four antigens. Studies in our normal and transgenic mice indicate that antigens targeting TbpB or TbpA can reduce colonization, thus the prospects of developing a vaccine that will eliminate bacteria expressing any of the four target antigens are promising.