Development of nonhuman primate model of S. aureus nasal carriage

金黄色葡萄球菌鼻携带非人灵长类动物模型的开发

• 批准号: 8953176
• 负责人: ALEXANDER MICHAEL COLE
• 金额: $ 26.14万
• 依托单位: UNIVERSITY OF CENTRAL FLORIDA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-16 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8953176
• 关键词: AIDS/HIV problem; Acute; Adhesives; Affect; Animal Model; Animals; Anterior nares; Anti-Bacterial Agents; Bacteria; Binding Proteins; Biological; Biology; Carrier State; Cells; Cessation of life; Chronic; Clinical; Clinical Trials; Communities; Development; Disease; Epithelial; Epithelial Cells; Experimental Models; Fibronectins; Genes; Genotype; Growth; Host Defense; Human; Human Activities; Immune response; In Vitro; Individual; Integration Host Factors; Interleukin-1; Investigation; Liquid substance; Macaca; Macaca nemestrina; Mediating; Modeling; Monkeys; Mucous Membrane; Mupirocin; Mus; Natural Immunity; Nose; Nosocomial Infections; Outcome; Participant; Phase; Primates; Proteomics; Recovery; Reporting; Research; Role; Sequence Homology; Staphylococcus aureus; Structure of mucous membrane of nose; Study models; Surface; Surveys; Testing; Tissue Model; Tissues; Washington; Work; antimicrobial; base; clinically relevant; cohort; cost effective; genetic analysis; in vivo; inflammatory marker; killings; microbial; model development; nonhuman primate; pathogen; polypeptide; public health relevance; response; safety testing

 DESCRIPTION (provided by applicant): Staphylococcus aureus (SA) nasal carriage is a common condition that predisposes individuals to severe community-acquired and nosocomial infections, and acts as an important reservoir for spreading pathogenic strains. We are beginning to understand that host and bacterial determinants collectively contribute to the carrier state, and have utilized nasal epithelial cells and ex vivo tissue models of the upper airways to better define the role of bacterial and host determinants of SA nasal carriage. However, to confirm the in vivo role of these factors in nasal carriage, a suitable animal model of SA colonization would be required. While useful, murine models are not ideal for studying longterm SA nasal carriage or the host's response to colonization, since these animals are not naturally colonized by SA, extraordinarily high inocula result only in transient colonization, and murine noses poorly model the mucosal host response of human noses. Thus, current models do not afford the ability to test SA carriage in its natural state. Nonhuman primates would likely better serve to model human SA nasal carriage. We explored whether pigtailed macaques could be used as a biologically relevant model of experimental SA nasal colonization, and discovered that over half were nasally colonized by SA. Genetic analyses revealed sequence homology between nasal SA of humans and macaques, suggesting that pigtailed macaque noses could be experimentally colonized with human-derived SA strains. This proposal seeks to develop a tractable, cost-effective model of nasal carriage of SA in pigtailed macaques, and utilize the model to assess the host's mucosal response to SA colonization. In Aim 1, we will develop the pigtailed macaque nasal carriage model using each macaque's own SA nasal isolate, and then use the model to explore how clinically relevant nasal human SA strains colonize macaque noses. Following experimental nasal colonization with macaque and clinically relevant nasal human SA strains, level of carriage will be assessed for two months by quantifying bacterial load and genotyping the recovered SA to confirm whether colonization persists with the inoculated strains. In Aim 2, we will utilize the pigtailed macaque model to determine how nasal carriage of SA affects innate mucosal host defense in vivo. Given we have reported that nasal fluids from human carriers of SA are deficient in killing SA in vitro, we will determine whether nasal fluids from macaques that carry SA in their noses are similarly deficient before and after in vivo experimental SA colonization. We will also utilize a robust proteomic approach to evaluate the biological effect of experimental SA colonization on the expression of antimicrobial polypeptides, inflammatory markers, and other mucosal host defense components. Collectively, we would provide the field with a nonhuman primate model for studying determinants of SA nasal carriage, and testing the safety and efficacy of candidate anti-SA nasal prophylaxes.

 描述（由申请人提供）：鼻腔携带金黄色葡萄球菌（SA）是一种常见疾病，使个体容易遭受严重的社区获得性和医院感染，并且是传播致病菌株的重要储存库。我们开始了解宿主和细菌决定因素共同影响携带者状态，并利用鼻上皮细胞和上呼吸道离体组织模型来更好地定义细菌和宿主决定因素在SA鼻携带中的作用。然而，为了证实这些因素在鼻运输中的体内作用，需要一种合适的动物模型 需要南澳殖民。虽然小鼠模型有用，但对于研究长期 SA 鼻腔携带或宿主对定植的反应并不理想，因为这些动物并非天然被 SA 定植，极高的接种量只会导致短暂的定植，而鼠类鼻子很难模拟人类鼻子的粘膜宿主反应。因此，当前模型无法在自然状态下测试 SA 承载。非人类灵长类动物可能会更好地模拟人类 SA 鼻腔运输。我们探讨了猪尾猕猴是否可以用作实验性 SA 鼻腔定植的生物学相关模型，并发现超过一半的尾猴鼻腔被 SA 定植。遗传分析揭示了人类和猕猴的鼻SA之间的序列同源性，这表明辫子猕猴的鼻子可以通过实验在人类衍生的SA品系上定植。 该提案旨在开发一种易于处理、经济有效的猪尾猕猴鼻腔携带SA的模型，并利用该模型评估宿主对SA定植的粘膜反应。在目标 1 中，我们将使用每只猕猴自身的 SA 鼻分离株开发猪尾猕猴鼻腔模型，然后使用该模型探索临床相关的人类鼻 SA 菌株如何在猕猴鼻子中定殖。在用猕猴和临床相关的人鼻 SA 菌株进行实验性鼻定植后，将通过量化细菌负荷和对回收的 SA 进行基因分型来评估两个月的携带水平，以确认接种菌株的定植是否持续存在。在目标 2 中，我们将利用尾猴模型来确定 SA 的鼻腔携带如何影响体内先天粘膜宿主防御。鉴于我们已报道人类SA携带者的鼻液在体外不足以杀死SA，我们将确定鼻中携带SA的猕猴的鼻液在体外实验前后是否同样缺乏杀灭SA的能力。 体内实验SA定植。我们还将利用强大的蛋白质组学方法来评估实验性 SA 定植对抗菌多肽、炎症标记物和其他粘膜宿主防御成分表达的生物学效应。总的来说，我们将为该领域提供一个非人类灵长类动物模型，用于研究 SA 鼻腔携带的决定因素，并测试候选抗 SA 鼻腔预防的安全性和有效性。