Host: Pathogen Interaction in Plague

主持人：鼠疫中病原体的相互作用

• 批准号: 8236988
• 负责人: Scott A Minnich
• 金额: $ 37.57万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8236988
• 关键词: Adherence; Adjuvant; Agonist; Animals; Antibiotic Therapy; Antibodies; Antigens; Applied Research; Area; Attenuated; Bacteria; Basic Science; Bioinformatics; Caenorhabditis elegans; Cell surface; Cells; Centers of Research Excellence; Charge; Communicable Diseases; Development; Dissection; Emerging Communicable Diseases; Extracellular Protein; Fleas; Funding; Genes; Genetic; Genome; Glucose; Goals; Human; Immune response; Immunity; Infection; Lead; Libraries; Lipid A; Maintenance; Methods; Modeling; Molecular; Mus; Mutation; Natural Immunity; Pathogenesis; Plague; Pneumonic Plague; Process; Production; Pseudogenes; Public Health; Rattus; Receptor Activation; Regimen; Regulation; Regulator Genes; Research; Resistance; Role; Screening procedure; Serum; Surface; TLR4 gene; Testing; Therapeutic; Transgenic Organisms; Vaccination; Vaccine Adjuvant; Vaccines; Virulence; Virulence Factors; Virulent; Yersinia; Yersinia pestis; adaptive immunity; bactericide; biodefense; cytokine; inorganic phosphate; mimetics; monolayer; mortality; nonhuman primate; novel; novel therapeutics; novel vaccines; pathogen; product development; protective effect; receptor; repaired; response; restoration; subcutaneous; success; therapeutic vaccine; toll-like receptor 4; vector

Our project previously focused on the development of novel vaccine and therapeutic strategies using Yersinia pestis models. We examined lipid A mimetics (aminoalkyl glucosaminide phosphates, AGPs), Tolllike receptor (TLR) 4 agonists. We found that AGPs directly protect against pneumonic plague, dramatically augment antibiotic therapy, and are efficacious adjuvants for either intranasal (i.n.) or subcutaneous (s.c.) vaccines containing Y. pestis capsular (F1) and/or V-antigens. Our vaccination regimen results in a rapid and sustained acquired TH1 immune response which protects in mouse and rat models. We also are characterizing several newly identified Y. pestis virulence factors and their regulation. Significant advances have been made toward identifying additional virulence mechanisms by screening for attenuated strains using advanced bioinformatic analyses, comprehensive transposon libraries, and through animal and Caenorhabditis elegans models. Our specific aims in this proposal expand on these successes: Specific Aim 1: To further examine the transition between induction of innate immunity with TLR agonists and specific immunity directed against pneumonic plague. The product development of AGPs requires delineation of the differences between murine and human immune responses. Transgenic humanized TLR4/MD-2 mice will be used to assess responses to AGPs as therapeutics and as vaccine adjuvants. In addition we will examine combinations of various TLR agonists to increase protection. Specific Aim 2: Determine the role of regulatory genes and surface/extracellular proteins in Y. pestis infection and/or flea vector maintenance. We will systematically characterize regulatory mutations, cell surface/extracellular proteins, and glucose-regulated genes for their role in virulence with the ultimate goal of identifying novel targets for product development. Specific Aim 3: Identify natural mutations (pseudogenes) that are essential for Y. pestis pathogenesis. We will test the hypothesis that increased bacterial virulence is, in part, due to evolutionary loss of genetic information during adaptation to the host. We will use bioinformatics to identify the lost and/or inactivated functions in the genomes of Y. pestis strains which are functional in the closely related but less virulent Yersinia enteropathogens. These identified Y. pestis 'pseudogenes' will be repaired to determine if this restoration attenuates virulence. We refer to this process as 'reverse molecular Koch's postulates'.

我们的项目以前专注于使用新型疫苗和使用治疗策略的开发 耶尔西尼亚瘟疫模型。我们检查了脂质A MIMETICS（氨基烷基葡萄糖胺磷酸盐，AGP），Tollike 受体（TLR）4激动剂。我们发现AGP直接保护肺鼠疫，急剧 增强抗生素治疗，是鼻内（I.N.）或皮下（S.C.）的有效佐剂 含有Y. Pestis囊囊（F1）和/或V-抗原的疫苗。我们的疫苗接种方案导致迅速 并持续获得的Th1免疫反应，可保护小鼠和大鼠模型。我们也是 表征了几个新鉴定的柴油毒力因子及其调节。重大进展 通过筛查减毒菌株来识别其他毒力机制 使用先进的生物信息学分析，全面的转座子库以及动物和 秀丽隐杆线虫模型。我们在此提案中的具体目标扩展了这些成功： 特定目的1：进一步研究与TLR先天免疫诱导之间的过渡 针对肺炎鼠疫的激动剂和特异性免疫力。 AGP的产品开发 需要描述鼠和人类免疫反应之间的差异。转基因 人源化TLR4/MD-2小鼠将用于评估对AGP的反应作为治疗剂和疫苗 佐剂。此外，我们将检查各种TLR激动剂的组合以增加保护。 特定目标2：确定调节基因和表面/细胞外蛋白在Y. Pestis中的作用 感染和/或跳蚤矢量维护。我们将系统地表征调节突变，细胞 表面/细胞外蛋白和葡萄糖调节的基因在毒力中的作用，最终目标是 确定新颖的产品开发目标。 特定目的3：识别对Y. Pestis必不可少的自然突变（假基因） 发病。我们将检验以下假设，即增加细菌毒力的部分是由于进化 适应宿主期间遗传信息的丧失。我们将使用生物信息学来识别丢失和/或 在密切相关但较少的尼氏菌菌株基因组中灭活功能 强烈的耶尔森氏病。这些确定的鼠疫杆菌的“假基因”将被修复，以确定是否是否 这种恢复会减轻毒力。我们将此过程称为“反向分子科赫的假设”。