Novel carbohydrate binding functions of the CR3 I-domain modulate gonococcal-cervical cell interactions

CR3 I 结构域的新型碳水化合物结合功能调节淋球菌-宫颈细胞相互作用

• 批准号: 10078936
• 负责人: Jennifer L Edwards
• 金额: $ 46.5万
• 依托单位: RESEARCH INST NATIONWIDE CHILDREN'S HOSP
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-16 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10078936
• 关键词: Adhesions; Affect; Agonist; Bacteria; Bacterial Proteins; Behavior; Binding; Binding Sites; Biological; C3bi; Calorimetry; Carbohydrates; Cell Communication; Cells; Centers for Disease Control and Prevention (U.S.); Cervical; Cervicitis; Cervix Uteri; Cessation of life; Coculture Techniques; Complement; Data; Development; Diagnosis; Disease; Ensure; Epithelial; Epithelial Cells; Event; Female; Fimbriae Proteins; Foundations; Frequencies; Future; Goals; Gonorrhea; HIV Infections; Health; Human; Immune; Immune response; Immune system; Immunity; Immunologic Receptors; In Vitro; Infection; Infertility; Innate Immune Response; Lectin; Ligands; Link; Literature; Macrophage-1 Antigen; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Medical Care Costs; Molecular; Morbidity - disease rate; Mucosal Immunity; Mucous Membrane; Multi-Drug Resistance; Natural Immunity; Nature; Neisseria gonorrhoeae; Outcome; Pathogenesis; Pathway interactions; Pattern recognition receptor; Pelvic Inflammatory Disease; Peptides; Phagocytes; Phase; Pilum; Play; Polysaccharides; Post-Translational Protein Processing; Prevalence; Prevention; Preventive measure; Process; Proteins; Public Health; Recombinants; Reporting; Role; Seminal; Signal Transduction; Structure; Surface; Surface Plasmon Resonance; Testing; Therapeutic; United States; Ursidae Family; Variant; Woman; Work; appendage; base; experience; glycosylation; human disease; human pathogen; improved; innate immune function; innovation; insight; macrophage; microbial; neutrophil; novel; novel strategies; novel therapeutics; pathogen; pathogenic bacteria; prevent; receptor; reproductive tract; response; sugar; trafficking; vaccine development

PROJECT SUMMARY The prevalence of Neisseria gonorrhoeae (Ng), its associated morbidity, and the emergence of untreatable strains, support a critical need for new preventative and therapeutic strategies for this important human- pathogen. Such developments require a complete understanding of the pathobiology of this archetypal, host- adapted pathogen. We show that phase-variable, glycan post-translational modifications on Ng surface appendages, pili (fimbriae), play a vital role in human infection. Complement receptor 3 (CR3) is an important pattern recognition receptor that is the key receptor mediating Ng colonization of human cervical mucosa. CR3 contains an “I-domain” region, which is known as a binding site for protein ligands. However, we found that Ng binding to the I-domain is mediated by the pilin-linked glycan (PLG). This is a seminal finding in innate immunity, as all previous literature ascribe CR3 carbohydrate binding to a separate, so called "lectin" domain. This was also the first demonstration that post-translational modifications made to a bacterial protein modulate pathogenesis. These findings will impact our understanding of microbial pathogenesis and innate immune responses. The goals of the present application are to define the specific contribution of the six, naturally occurring, PLG structures in mediating the Ng-CR3 I-domain interaction and to determine the biological relevance of each of these interactions to infection in females. Guided by strong preliminary data, we hypothesize that variable PLG structures initiate key, but highly divergent, outcomes with CR3 I-domain engagement. We will resolve our hypothesis through two specific aims: Aim 1) Define the effect of variation in pilin glycan structure on direct PLG-I-domain interactions; we will define the specific molecular interactions occurring with the CR3 I-domain for the six natural Ng PLG structures. Aim 2) Define the effect of PLG-I- domain interactions on Ng pathogenesis and CR3 function; we will define targeted epithelial cell responses to CR3 I-domain engagement by Ng that bear different PLG structures and their effect on Ng pathogenesis. Our approach is innovative in using biologically relevant human primary cervical cells, both alone and in co-culture with phagocytes, combined with low passage Ng isolates. This ensures that data obtained are relevant to human processes. By defining the effector functions controlled by I-domain lectin activity on primary human epithelial cells, we will provide critical new information regarding Ng pathogenesis, CR3 function, and the fundamental mechanisms that govern human cervical mucosal immunity. Moreover, we will define the utility of targeting the PLG-CR3 I-domain interaction as a new, and improved, host-targeted approach to treat and/or prevent Ng disease in women. Our studies will very likely impact work on other human pathogens and on innate immunity, broadening the significance of our outcomes. We have extensive experience in defining the molecular mechanisms of Ng pathogenesis. We are ideally placed to do this work in having discovered CR3 as the key receptor for Ng cervical infection and the biosynthetic pathways for pili glycosylation.

项目摘要 Neiserseria Gonorrhoeae（NG）的流行，其相关的发病率和不可治疗的出现 应变，支持对这一重要人类的新预防和治疗策略的迫切需求 病原体。 适应的病原体。 附属物，pili（fimbriae），在人类感染中起着至关重要的作用。 模式识别受体是介导人宫颈粘膜NG定殖的关键受体。 包含一个“ i域”区域，我们被称为蛋白质配体的结合位点 NG与I域的结合是由pilin lineked glucan（PLG）介导的。 免疫力，如以前所有文献cr3碳水化合物与单独的所谓“凝集素”结构域结合。 这也是第一次证明对细菌蛋白进行的翻译 调节发病机理。 免疫反应。 自然发生，介导Ng-Cr3 I域相互作用并确定您的PLG结构 每种相互作用与女性感染的相关性。 假设可变的PLG结构被构成关键，但具有高度分歧的CR3 I域的结果 参与。我们将通过两个特定目的解决我们的假设： 直接PLG-1域相互作用的pilin聚糖结构；我们将定义特定的分子相互作用 用于六个天然NG PLG结构的CR3 I域。 NG发病机理和CR3功能的域相互作用； CR3 i-domain通过NG的参与度具有不同的PLG结构及其对NG发病机理的影响 方法是使用与生物学相关的人类原代宫颈细胞的创新性，无论是单独还是共同培养 使用吞噬细胞，结合通道NG分离株。 人类过程。 上皮细胞，我们将提供有关NG发病机理，CR3功能和TEE的关键新信息 控制人类宫颈粘膜免疫的基本机制。 针对PLG-CR3 I域I-Domain I-Domain International的目标是新的，改进的宿主目标方法和/或 预防女性的NG疾病可能会影响其他人类病原体 先天免疫，扩大结果的重要性。 NG发病机理的分子机制。 NG宫颈感染的关键受体和Pili糖基化的生物合成途径。