Targeting Pneumococcal Colonization

针对肺炎球菌定植

• 批准号: 10113534
• 负责人: Jeffrey Neal Weiser
• 金额: $ 21.19万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10113534
• 关键词: Address; Adjuvant; Adult; Affect; Age; Anabolism; Animals; Antibodies; Antibody Response; Antigen Targeting; Antigens; Biology; Cell Wall; Cell surface; Child; Data Set; Development; Disease; Enzymes; Extracellular Protein; Fibrinogen; Formulation; Future; Genes; Genetic Screening; Genome; Glycoconjugates; Growth; Human; Immune; Immunity; Immunization; Immunize; Infant; Infection; Interruption; Laboratories; Licensing; Maintenance; Metabolism; Modeling; Mucosal Immunity; Mucous Membrane; Mus; Mutagenesis; N-acetylglucosamine deacetylase; Neuraminidase; Organism; Pathogenesis; Penicillin-Binding Proteins; Peptide Hydrolases; Phosphorylcholine; Pneumococcal Colonization; Pneumococcal Infections; Polysaccharides; Population; Prevalence; Prevention; Prevention strategy; Protein Kinase; Proteins; Public Health; Recombinant Proteins; Recombinants; Respiratory System; Serotyping; Source; Streptococcus pneumoniae; Surface; Surface Antigens; Technology; Teichoic Acids; Testing; Upper respiratory tract; Vaccination; Vaccines; Variant; base; contagion; cross reactivity; esterase; experience; immunogenic; in vitro Assay; member; mouse model; murine antibody; mutant; novel; novel strategies; pressure; prevent; protein function; pup; transmission process; transposon sequencing; vaccine development; vaccine discovery

Streptococcus pneumoniae (Spn, the pneumococcus) remains a leading cause of respiratory tract and invasive infection. While vaccination with capsular polysaccharide-based vaccines has decreased the high burden of Spn disease, this approach fails to target the majority of Spn, which are non-vaccine serotypes. Unfortunately, the development of more broadly-acting, serotype- independent vaccines that focus on preventing Spn disease has proven to be elusive. The required first step for Spn-host interaction, however, is colonization of the mucosal surfaces of the upper airways. Our premise is that interrupting colonization will have the greatest overall impact on Spn disease and offers new, unexplored possibilities for prevention. We have optimized an infant mouse model of Spn colonization that has allowed the application of Tn-Seq transposon mutagenesis to interrogate the entire Spn genome to identify loci affecting colonization. The complete dataset of non-essential Spn genes affecting colonization was then compared to bacterial surface factors that are immunogenic during human infection. Defined mutants in each of these candidates were constructed and tested in both infant and adult murine models of colonization. Seven loci expressing proteins of known function that modify the bacterial surface or host substrates were validated as required for efficient colonization. These proteins are all members of the Spn core genome present in all strains and show minimal sequence variation across isolates. Our hypothesis is that immunity to these candidates, alone or in combination, will interrupt Spn colonization. This hypothesis will be tested first by immunization with recombinant protein in adjuvant (SC or IN) to maximize antibody responses. Murine antibody will be used to confirm surface localization and to explore its ability to block protein function using in vitro assays. Immune mice v. adjuvant alone controls will then be challenged IN or via pup-to-pup transmission to assess protection from colonization with diverse strains. In future studies, candidates validated in the murine model can be investigated using experimental human pneumococcal carriage to bridge the critical step from animal to human testing. Thus, our proposal takes a novel approach, targeting colonization, to the ongoing public health problem of the pneumococcus.

肺炎链球菌（SPN，肺炎球菌）仍然是呼吸道的主要原因 和侵入性感染。虽然基于囊状多糖疫苗接种疫苗的疫苗已有 SPN疾病的高负担减轻了，这种方法无法针对大多数SPN，而SPN 是非疫苗血清型。不幸的是，发展更广泛，血清型的发展 专注于预防SPN疾病的独立疫苗被证明是难以捉摸的。这 但是 上呼吸道。我们的前提是中断殖民化的总体总体最大 对SPN疾病的影响，并为预防提供了新的，未开发的可能性。我们已经优化了 SPN定植的婴儿小鼠模型，该模型允许应用TN-Seq Transposon 诱变以询问整个SPN基因组，以鉴定影响定植的基因座。这 然后将影响定植的非必需的SPN基因的完整数据集进行比较 人类感染过程中免疫原性的细菌表面因子。每个定义突变体 在婴儿和成人鼠模型中构建和测试了这些候选者中的 殖民化。七个基因座表达已知功能的蛋白质，可修饰细菌表面 或根据需要进行有效定植的需要验证宿主底物。这些蛋白质都是 SPN核心基因组的成员存在于各种菌株中，并显示最小的序列变化 跨分离株。我们的假设是，对这些候选人的免疫力单独或结合了 中断SPN定植。该假设将首先通过重组免疫进行检验 辅助（SC或ON）中的蛋白质，以最大化抗体反应。鼠抗体将用于 确认表面定位并探索其使用体外测定法阻断蛋白质功能的能力。 免疫小鼠诉单独的佐剂对照将在幼崽到幼崽传播中受到挑战 评估不同菌株的保护免受定殖。在未来的研究中，候选人得到了验证 在鼠模型中，可以使用实验性人肺炎球菌进行研究 桥梁从动物到人类测试的关键步骤。因此，我们的建议采取了一种新颖的方法， 靶向殖民化，涉及肺炎球菌的持续公共卫生问题。