Rational design of transferrin binding protein-based vaccines to combat gonorrhea

合理设计基于转铁蛋白结合蛋白的淋病疫苗

• 批准号: 9888316
• 负责人: CYNTHIA N CORNELISSEN
• 金额: $ 62.67万
• 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: 2019-03-06 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9888316
• 关键词: Adjuvant; Animal Model; Antibiotic Resistance; Antigenic Variation; Antigens; Antimicrobial Resistance; Binding; Cellular Immunity; Chimera organism; Crystallization; Crystallography; Data; Development; Disease; Disease model; Dose; Engineering; Epitopes; Exhibits; Female; Frequencies; Gender; Goals; Gonorrhea; Hemoglobin; Human; Humoral Immunities; Immune response; Immunity; Immunization; Immunologic Memory; Immunologics; Infection; Inflammation; Iron; Iron-Binding Proteins; Lactoferrin; Lead; Ligand Binding; Ligands; Lipoproteins; Maps; Mediator of activation protein; Membrane Transport Proteins; Methods; Modeling; Mucosal Immunity; Multi-Drug Resistance; Mutagenesis; Neisseria gonorrhoeae; Neisseria meningitidis; Nutrient; Nutritional Immunity; Pathogenicity; Pelvic Inflammatory Disease; Preventive vaccine; Process; Protein Conformation; Proteins; Publishing; Resistance development; Role; Route; Schedule; Self Tolerance; Sexually Transmitted Diseases; Siderophores; Source; Structure; Superbug; Surface; Syndrome; System; TFRC gene; Testing; Therapeutic; Tissues; Transferrin; Transferrin-Binding Proteins; Urethritis; Uterus; Vaccination; Vaccine Antigen; Vaccine Design; Vaccines; Variant; base; combat; cross reactivity; design; experimental study; human pathogen; humanized mouse; immunogenicity; in silico; in vivo; innovation; male; mouse model; mucosal site; mutant; novel; pathogen; protective efficacy; reproductive tract; vaccine candidate; vaccine delivery; vaccine development; volunteer

The human pathogen, Neisseria gonorrhoeae (Ngo) is capable of utilizing human transferrin as the sole source of iron. Efficient iron transport from human transferrin (hTf) requires expression of a TonB-dependent, integral outer membrane transporter (TbpA), and a surface-exposed lipoprotein (TbpB). Expression of the Tbps is necessary for the gonococcus to establish human infection in male volunteer studies, implying a critical role for these proteins in the human host. Because this transport system is expressed by all gonococci and the protein components are well conserved, their potential as vaccine targets will be explored in this proposal. The overarching hypothesis to be tested in the proposed study is that a rationally-designed vaccine consisting of engineered TbpA and TbpB antigens, combined with an experimentally selected adjuvant, will provide immunologic cross-protection against both asymptomatic colonization and pathogenic inflammation caused by diverse N. gonorrhoeae strains. The specific aims of the proposal are as follows: Aim 1. Rational design of Tbp-based vaccines. In this aim, structures of the gonococcal Tbps will be defined with and without hTf. These structures will guide mutagenesis efforts to produce vaccine antigens defective in ligand binding and lacking hypervariable sequences with minimal structure disruption. Aim 2. Optimizing vaccine delivery in humanized female mouse model of lower genital tract infection. Adjuvants, delivery routes, doses and schedules will be tested for optimal protection in the female lower genital tract infection model using humanized mice expressing hTf. Immunological correlates and determinants of protection will be defined by broadly testing humoral and cellular immunity factors. Aim 3. Test for protective efficacy of Tbp-based vaccines in new humanized infection models. Optimized antigens, adjuvants, routes and schedules will be tested for protection in humanized mouse models of pelvic inflammatory disease, male urethritis and nasopharyngeal infection. Again, immunological correlates and determinants of protection in these new animal models of infection will be characterized. Aim 4. Evaluate rationally designed vaccines for cross protection in all models of infection. Optimized, rationally- designed vaccine formulations will be tested for protection against a broad group of Ngo strains, including antimicrobial resistant “superbug” strains. Immunological correlates of protection will be validated with these strains in all of the humanized mouse models of infection. These studies are significant since they may lead to the development of an efficacious vaccine against a recalcitrant pathogen that has developed resistance to existing therapeutic methods. These studies are innovative because they will employ structure-guided vaccine design to develop ligand-binding incompetent vaccine antigens, which are hypothesized exhibit enhanced immunogenicity compared to the wild-type proteins. Moreover, these antigens will be tested for protection in novel humanized mouse models of infection and immunological correlates of protection will be defined, closing a key gap in our understanding of immunity against Ngo.

人类病原体淋病奈瑟菌 (Ngo) 能够利用人转铁蛋白作为唯一来源 人转铁蛋白 (hTf) 的有效铁转运需要依赖 TonB 的表达。 外膜转运蛋白（TbpA）和表面暴露的脂蛋白（TbpB）的表达是。 在男性志愿者研究中，淋球菌是建立人类感染所必需的，这意味着淋球菌的关键作用 因为这种转运系统是由所有淋球菌和蛋白质表达的。 成分得到了很好的保存，本提案将探讨它们作为疫苗靶标的潜力。 在拟议的研究中要测试的总体假设是，合理设计的疫苗包括 工程化的 TbpA 和 TbpB 抗原与实验选择的佐剂相结合，将提供 针对无症状定植和由以下原因引起的致病性炎症的免疫交叉保护 本提案的具体目标如下： 目的 1. 合理设计淋病奈瑟菌菌株。 基于 Tbp 的疫苗将在使用和不使用 hTf 的情况下定义淋球菌 Tbp 的结构。 结构将指导诱变工作，以产生配体结合缺陷和缺乏的疫苗抗原 目标 2. 优化人源化疫苗的递送。 雌性小鼠下生殖道感染模型的佐剂、给药途径、剂量和时间表。 使用表达的人源化小鼠在女性下生殖道感染模型中测试了最佳保护 hTf. 免疫学相关性和保护的决定因素将通过广泛测试体液和 目标3.测试基于Tbp的疫苗对新的人源化感染的保护效力。 优化的抗原、佐剂、途径和时间表将在人源化小鼠中进行保护测试。 盆腔炎、男性尿道炎和鼻咽感染模型。 目标 4 将描述这些新的感染动物模型中保护的相关因素和决定因素。 评估合理设计的疫苗在所有感染模型中的交叉保护作用。 设计的疫苗配方将进行针对广泛的非政府组织菌株的保护测试，包括 抗菌素耐药性“超级细菌”菌株的保护作用的免疫学相关性将通过这些进行验证。 这些研究很重要，因为它们可能导致所有人源化小鼠感染模型中的菌株。 开发针对顽固病原体的有效疫苗，该病原体已产生抗药性 这些研究具有创新性，因为它们将采用结构引导的疫苗。 设计开发配体结合无能疫苗抗原，其被利用表现出增强 此外，将测试这些抗原的保护作用。 将定义新型人源化小鼠感染模型和保护的免疫相关性，结束 我们对非政府组织豁免理解的一个关键差距。