Project-002

项目-002

• 批准号: 10330127
• 负责人: CYNTHIA N CORNELISSEN
• 金额: $ 10.05万
• 依托单位: GEORGIA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-25 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10330127
• 关键词: Acidic Amino Acids; Acidic Region; Antibiotics; Antigen Targeting; Antigens; Bacteria; Binding; Binding Proteins; Cations; Cervical; Charge; Detergents; Development; Dimensions; Disease; Disease model; Engineering; Epitopes; Etiology; Excision; Female genitalia; Formulation; Funding; Goals; Gonorrhea; Grant; Human; Hybrids; Immune response; Immunoassay; Immunologics; Individual; Infection; Infection prevention; Iron; Iron-Binding Proteins; Lactoferrin; Libraries; Lipoproteins; Lobe; Mediating; Membrane; Membrane Transport Proteins; Modeling; Mutation; Neisseria gonorrhoeae; Nutrient; Pathway interactions; Pelvic Inflammatory Disease; Peptides; Production; Property; Proteins; Recombinants; Resistance; Role; Sexually Transmitted Diseases; Solubility; Structure; Surface; System; TFRC gene; Testing; Transferrin; Transferrin-Binding Protein A; Transferrin-Binding Protein B; Transgenic Mice; United States National Institutes of Health; Upper respiratory tract; Urethra; Uterus; Vaccinated; Vaccine Design; Vaccines; Variant; antigen test; base; cross reactivity; design; human male; insight; lactoferrin receptors; male; mouse model; mutant; novel; pathogen; periplasm; programs; protein B; receptor; reproductive tract; response; scaffold; scale up; urogenital tract; vaccine development; vaccine evaluation

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.

该项目的目标是开发一种针对淋病奈瑟菌（淋病奈瑟菌）的广泛交叉保护性疫苗。 导致常见性传播疾病淋病的细菌。 N菌株的出现。 对大多数抗生素具有抗药性的淋病引起了人们对淋病无法治愈的担忧，并补充道 迫切需要开发和实施有效的疫苗来预防这种感染及其后果 后遗症。淋病奈瑟菌是人类特有的病原体，仅存在于泌尿生殖道或更少 通常发生在人类宿主的上呼吸道。我们的方法是针对表面组件 负责介导从宿主铁结合蛋白、转铁蛋白 (Tf) 和乳铁蛋白 (Lf) 中获取铁， 因为这两个系统已被证明对于细菌的生存及其引起的能力至关重要 人类男性尿道感染模型中的感染。由于没有这些受体细菌就无法生存 蛋白质，它们将无法通过受体的丧失而逃脱疫苗的覆盖，因此可以被消除 如果我们成功地开发出针对所有人的完全交叉保护性疫苗，则来自已接种疫苗的个体 受体蛋白的变体。 该项目的重点是使用结构引导的抗原设计来生成靶向的工程抗原 表面脂蛋白、乳铁蛋白结合蛋白 B (LbpB) 和完整的外膜转运 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 针对这些蛋白质的所有已知变体的反应。我们新颖的综合疫苗设计和评估 管道方法将在免疫测定中使用抗原变体库，在定植中使用菌株库 和感染模型来评估免疫反应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，这些突变体是 预计将共同提供针对所有 LbpB 变体的广泛交叉保护性免疫反应。我们将 评估 LbpB C 叶带负电区域的免疫学特性，确定 去除它们的影响，并确定可在样品上展示的 LbpA 表位的最佳组合 LbpB C 叶可诱导针对 LbpA 的广泛交叉保护反应。 最后一步是整合 NIH 已设立的专注于开发抗原的拨款的结果 靶向转铁蛋白结合蛋白 B 和 A（TbpB 和 TbpA）并确定最佳组合 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 针对所有四种抗原的免疫反应。对正常小鼠和转基因小鼠的研究表明 针对 TbpB 或 TbpA 的抗原可以减少定植，因此开发疫苗的前景 消除表达四种靶抗原中任何一种的细菌是有希望的。