Development of nonhuman primate model of S. aureus nasal carriage

 描述（由申请人提供）：金黄色葡萄球菌（SA）鼻携带是一种常见的疾病，使个体易于发生严重的社区获得性和医院感染，并作为传播致病菌株的重要储存库。我们开始了解到，主机和细菌的决定因素共同有助于运营商的状态，并利用鼻上皮细胞和离体组织模型的上呼吸道，以更好地定义SA鼻腔运输的细菌和主机的决定因素的作用。然而，为了证实这些因子在鼻携带中的体内作用，需要一种合适的动物模型， 殖民地化是必须的。虽然有用，但鼠模型对于研究长期SA鼻携带或宿主对定殖的反应并不理想，因为这些动物不是天然地被SA定殖，非常高的接种物仅导致短暂的定殖，并且鼠鼻子对人鼻子的粘膜宿主反应的模拟很差。因此，目前的模型不具备在自然状态下测试SA运载的能力。非人灵长类动物可能更适合模拟人类SA鼻携带。我们探讨了是否可以使用猪尾猕猴作为实验SA鼻腔定植的生物学相关模型，并发现超过一半的SA鼻腔定植。遗传分析揭示了人类和猕猴的鼻SA之间的序列同源性，表明猪尾猕猴的鼻子可以通过实验与人源性SA菌株定殖。 该建议旨在开发一种易于处理的，具有成本效益的模型，在猪尾猕猴的鼻腔运输SA，并利用该模型来评估主机的粘膜响应SA殖民。在目标1中，我们将使用每只猕猴自己的SA鼻分离物开发猪尾猕猴鼻携带模型，然后使用该模型探索临床相关的鼻人类SA菌株如何定殖猕猴鼻子。在用猕猴和临床相关的人鼻SA菌株进行实验性鼻定殖后，将通过定量细菌载量和对回收的SA进行基因分型来评估携带水平两个月，以确认接种菌株是否持续定殖。在目标2中，我们将利用猪尾猕猴模型来确定SA的鼻携带如何影响体内固有粘膜宿主防御。鉴于我们已经报道了来自SA携带者的鼻液在体外杀死SA方面存在缺陷，我们将确定来自鼻中携带SA的猕猴的鼻液在体外杀死SA之前和之后是否存在类似的缺陷。 体内实验SA定殖。我们还将利用一个强大的蛋白质组学方法来评估实验SA殖民抗菌多肽，炎症标志物，和其他粘膜宿主防御组件的表达的生物学效应。总的来说，我们将为该领域提供一个非人灵长类动物模型，用于研究SA鼻携带的决定因素，并测试候选抗SA鼻粘膜松弛剂的安全性和有效性。

 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.