Development of an ex vivo porcine lung model for studying growth, virulence, and signaling of Pseudomonas aeruginosa.
Development of an ex vivo porcine lung model for studying growth, virulence, and signaling of Pseudomonas aeruginosa.
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DOI：
10.1128/iai.01554-14
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发表时间：
2014-08
影响因子：
3.1
通讯作者：
Diggle SP
中科院分区：
文献类型：
--
作者：
Harrison F;Muruli A;Higgins S;Diggle SP
对细菌病原体（例如铜绿假单胞菌）的慢性感染的研究使用各种体外和活宿主模型。虽然这些模型增加了我们对病原体生长、毒力和进化的理解，但每种模型都有一定的局限性。体外模型无法概括宿主器官的复杂空间结构，而活体宿主的实验由于伦理原因在样本量和感染持续时间方面受到限制;活体哺乳动物模型还需要运行成本高昂的专门设施。为了解决这个问题，我们开发了一种体外猪肺（EVPL）模型，用于在模拟慢性感染的囊性纤维化（CF）肺的环境中定量铜绿假单胞菌生长、群体感应（QS）、毒力因子产生和组织损伤。在我们的模型的第一个测试中，我们表明，lasR突变体，不响应3-氧代-C12-高丝氨酸内酯（HSL）介导的QS，表现出减少EVPL的毒力因子的生产。我们还表明，lasR突变体生长以及或更好地比相应的野生型菌株在EVPL。lasR突变体在慢性CF肺部感染期间频繁且反复出现，但控制其外观和传播的进化力量尚不清楚。我们的数据与lasR突变体在肺中充当社会“骗子”的假设不一致;相反，我们的结果支持lasR突变体更适应肺环境的假设。更一般地说，这种模型将有助于改善微生物疾病的研究，特别是研究相同和不同物种的细胞如何在空间结构化环境中的多微生物感染中相互作用。
Research into chronic infection by bacterial pathogens, such as Pseudomonas aeruginosa, uses various in vitro and live host models. While these have increased our understanding of pathogen growth, virulence, and evolution, each model has certain limitations. In vitro models cannot recapitulate the complex spatial structure of host organs, while experiments on live hosts are limited in terms of sample size and infection duration for ethical reasons; live mammal models also require specialized facilities which are costly to run. To address this, we have developed an ex vivo pig lung (EVPL) model for quantifying Pseudomonas aeruginosa growth, quorum sensing (QS), virulence factor production, and tissue damage in an environment that mimics a chronically infected cystic fibrosis (CF) lung. In a first test of our model, we show that lasR mutants, which do not respond to 3-oxo-C12-homoserine lactone (HSL)-mediated QS, exhibit reduced virulence factor production in EVPL. We also show that lasR mutants grow as well as or better than a corresponding wild-type strain in EVPL. lasR mutants frequently and repeatedly arise during chronic CF lung infection, but the evolutionary forces governing their appearance and spread are not clear. Our data are not consistent with the hypothesis that lasR mutants act as social “cheats” in the lung; rather, our results support the hypothesis that lasR mutants are more adapted to the lung environment. More generally, this model will facilitate improved studies of microbial disease, especially studies of how cells of the same and different species interact in polymicrobial infections in a spatially structured environment.