Harnessing Antibody Responses to Prevent and Treat Urinary Tract Infections

利用抗体反应预防和治疗尿路感染

• 批准号: 10523527
• 负责人: Andrew Leon Kau
• 金额: $ 62.77万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-11-10 至 2026-10-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10523527
• 关键词: Activities of Daily Living; Address; Adherence; Affect; Affinity; Animal Model; Animals; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Antibodies; Antibody Formation; Antibody Response; Antibody Specificity; Area; B-Lymphocytes; Bacterial Adhesins; Binding; Bladder; Blocking Antibodies; Cell Separation; Cells; Coal; Communicable Diseases; Communities; Cystitis; Data; Development; Diagnostic; Distal; Epithelium; Epitopes; Equilibrium; Escherichia coli; Extended-spectrum β-lactamase; Gastrointestinal tract structure; Generations; Genome; Gnotobiotic; Host Defense Mechanism; Human; Immune; Immune System Diseases; Immunization; Immunoglobulin G; Immunoglobulin-Secreting Cells; Impairment; Infection prevention; Invaded; Light; Mannose; Mannosides; Mediating; Modeling; Molecular Conformation; Monkeys; Monoclonal Antibodies; Mucous Membrane; Multi-Drug Resistance; Mus; Mutation; Operon; Oral Administration; Passive Immunization; Pathogenesis; Pathogenicity; Patients; Persons; Phase Ia/Ib Clinical Trial; Pilum; Prevalence; Prevention; Production; Public Health; Recombinants; Recurrence; Research; Resistance; Resolution; Role; Serinus; Signal Transduction; Specificity; Surface; Tertiary Protein Structure; Testing; Therapeutic; Therapeutic Uses; Thinking; Time; United States; Urethra; Urinary tract; Urinary tract infection; Urine; Uropathogen; Uropathogenic E. coli; Vaccinated; Vaccination; Vaccines; Virus Diseases; Woman; ascending urinary tract infection; bacterial community; cancer therapy; combat; design; effectiveness testing; fluoroquinolone resistance; gut colonization; gut microbiota; human monoclonal antibodies; humanized monoclonal antibodies; in vivo; microbial community; mouse model; mucosal vaccination; prevent; receptor; resistant strain; response

SUMMARY Urinary tract infections (UTIs) affect 15 million women in the United States every year and treatment for UTIs is becoming more difficult due to high rates of antibiotic resistance. Further, UTIs are highly recurrent. Between 20 and 40% of UTI episodes are followed by recurrent UTIs (rUTIs), with some women suffering as many as 6 or more recurrences per year. Uropathogenic Escherichia coli (UPEC) is the major causative agent of UTIs. Antibiotic resistance within UPEC isolates is rising, and the emergence of extended-spectrum beta-lactamase producing and fluoroquinolone resistant strains is a serious public health concern. Type 1 pili, tipped with the mannose binding FimH adhesin have been shown to be essential for bladder colonization and UTI pathogenesis in multiple mouse models. FimH mediates binding to mannosylated uroplakins lining the bladder surface to facilitate colonization and invasion into bladder cells where they rapidly replicate into intracellular bacterial communities that protect UPEC from immune cells and antibiotics. In addition, FimH facilitates the ability of UPEC to establish a reservoir in the GIT, from where they can seed UTIs by ascending from the periurethral area into the bladder. While UPEC are genetically variable, FimH is part of the core E. coli genome, although rare strains have been found with mutations in the type 1 operon. Immunization against FimH protects against UPEC UTI in murine and monkey cystitis models and a FimH-based vaccine has been allowed by the FDA for patients suffering from multi-drug resistant UPEC. In animal models, protection is antibody-mediated, as FimH- specific IgG antibodies are found in the urine from protected animals and can protect from UTI through passive transfer. Intriguingly, UPEC abundance in the gut is increased at the time of symptomatic UTI, suggesting that gut colonization is a key step in the rUTI cycle. Additionally, studies in this proposal show that eliciting a mucosal antibody response against FimH can reduce UPEC colonization of the gut. In light of these findings, this proposal addresses the hypothesis that anti-FimH induced antibodies can combat rUTI by two distinct mechanisms: i) prevention of UPEC binding to the uroepithelium; and ii) interference with UPEC colonization of the GIT, thereby lowering the likelihood of UPEC introduction into the urinary tract. The aims of this proposal are to: i) determine how mucosal vaccination against FimCH reduces UPEC gut colonization (Aim 1); ii) exploit vaccine induced B cell responses to isolate monoclonal antibodies (mAbs) to FimH and determine their epitope specificity (Aim 2); and iii) use these mAbs in mouse models of GIT colonization and cystitis in order to elucidate mechanisms of protection (Aim 3). The research plan will unravel the mechanisms by which anti-FimH antibodies may function to prevent UTIs by directly blocking bladder binding and indirectly by interfering with UPEC GIT colonization. These results will inform the rational targeting of the uropathogens that affect millions of people with rUTIs. In addition, our approach enables the rapid generation of anti-UPEC human mAbs that can be used for therapeutics and diagnostics.

概括 尿路感染（UTI）每年都会影响美国1500万妇女，而对UTI的治疗是 由于抗生素耐药性高，变得更加困难。此外，UTI是高度反复的。在20之间 40％的UTI发作之后是经常性UTI（RUTIS），一些妇女遭受了多达6个或6个或 每年更多的复发。尿道病大肠杆菌（UPEC）是UTI的主要病因。 UPEC分离株内的抗生素抗性正在上升，并且出现了延长的频谱β-内酰胺酶 产生和氟喹诺酮抗性菌株是一个严重的公共健康问题。 1型pili，带有 甘露糖结合FIMH粘附素已被证明对于膀胱定殖和UTI发病机理至关重要 在多个鼠标模型中。 FIMH介导与膀胱表面衬里的甘露糖基化的尿素结合到 促进定植并侵袭膀胱细胞，在那里它们迅速复制成细胞内细菌 保护UPEC免疫细胞和抗生素的社区。另外，FIMH促进了 UPEC在GIT中建立一个储层，从那里可以从尿道上升起utis。 区域进入膀胱。尽管UPEC在遗传上是可变的，但FIMH是核心大肠杆菌基因组的一部分，尽管 在1型操纵子中发现了罕见的菌株。防止FIMH的免疫预防 FDA允许使用Murine和Monkey Cystitis模型的UPI UTI以及基于FIMH的疫苗 患有多药耐药UPEC的患者。在动物模型中，保护是抗体介导的，如FIMH- 特定的IgG抗体在受保护动物的尿液中发现，可以通过被动保护UTI 转移。有趣的是，肠道中的UPEC丰度在有症状的UTI时增加，这表明 肠道定殖是RUTI周期中的关键步骤。此外，该提案中的研究表明，引起粘膜 对FIMH的抗体反应可以降低肠道的UPEC定植。鉴于这些发现，该提议 解决了以下假设：抗FIMH诱导的抗体可以通过两种不同的机制来对抗ruti：i） 预防UPEC结合到巨大上皮； ii）干扰git的UPEC殖民化 降低UPEC引入尿路的可能性。该提议的目的是：i）确定 针对FIMCH的粘膜疫苗接种如何减少UPEC肠道定殖（AIM 1）； ii）利用疫苗诱导的B 细胞对分离单克隆抗体（mAb）对FIMH的反应并确定其表位特异性（AIM 2）； iii）在GIT定植和膀胱炎的小鼠模型中使用这些mAB，以阐明 保护（目标3）。研究计划将揭示抗FIMH抗体可能起作用的机制 通过直接阻断膀胱结合并间接干扰UPEC GIT定殖，以防止UTI。 这些结果将为影响数百万鲁蒂人患者的尿道病的合理靶向。在 此外，我们的方法使可用于治疗剂的快速产生的反抗EC人物mAB 和诊断。