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Pseudomonas aeruginosa Infection of Zebrafish Involves both Host and Pathogen Determinants

机译：斑马鱼的铜绿假单胞菌感染涉及宿主和病原体的决定因素。

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Zebrafish (Danio rerio) have a number of strengths as a host model for infection, including genetic tractability, a vertebrate immune system similar to that of mammals, ease and scale of laboratory handling, which allows analysis with reasonable throughput, and transparency, which facilitates visualization of the infection. With these advantages in mind, we examined whether zebrafish could be used to study Pseudomonas aeruginosa pathogenesis and found that infection of zebrafish embryos with live P. aeruginosa (PA14 or PAO1) by microinjection results in embryonic death, unlike infection with Escherichia coli or heat-killed P. aeruginosa, which has no effect. Similar to studies with mice, P. aeruginosa mutants deficient in type three secretion (pscD) or quorum sensing (lasR and mvfR) are attenuated in zebrafish embryos infected at 50 h postfertilization (hpf), a developmental stage when both macrophages and neutrophils are present. In contrast, embryos infected at 28 hpf, when only macrophages are initially present, succumb to lethal challenge with far fewer P. aeruginosa cells than those required for embryos infected at 50 hpf, are susceptible to infection with lasR and pscD deletion mutants, and are moderately resistant to infection with an mvfR mutant. Finally, we show that we can control the outcome of infection through the use of morpholinos, which allow us to shift immune cell numbers, or small molecules (antibiotics), which rescue embryos from lethal challenge. Thus, zebrafish are a novel host model that is well suited for studying the interactions among individual pathogenic functions of P. aeruginosa, the role of individual components of host immune defense, and small-molecule modulators of infection.
机译：斑马鱼（ Danio rerio ）作为感染宿主模型具有许多优势，包括遗传易处理性，类似于哺乳动物的脊椎动物免疫系统，实验室操作的简便性和规模，这使得合理的分析成为可能。通量和透明性，可促进感染的可视化。考虑到这些优点，我们研究了斑马鱼是否可用于研究铜绿假单胞菌的发病机理，并发现斑马鱼胚胎被活的 P感染。与大肠杆菌或热灭活的 P感染不同，通过显微注射的铜绿假单胞菌（PA14或PAO1）导致胚胎死亡。铜绿，没有效果。与小鼠研究相似， P。在感染的斑马鱼胚胎中，缺乏三型分泌（ pscD ）或群体感应（ lasR 和 mvfR ）的铜绿突变体被减弱受精后50小时（hpf），这是同时存在巨噬细胞和中性粒细胞的发育阶段。相反，当最初仅存在巨噬细胞时，以28 hpf感染的胚胎因极少的 P而死于致命的攻击。绿脓杆菌细胞比感染50 hpf的胚胎所需的细胞要多，它们容易感染 lasR 和 pscD 缺失突变体，并且对< em> mvfR 突变体。最后，我们证明了我们可以通过使用吗啉代菌来控制感染的结果，这使我们能够转移免疫细胞数量或小分子（抗生素），从而使胚胎免于致命的攻击。因此，斑马鱼是一种新型宿主模型，非常适合研究 P的各个致病功能之间的相互作用。铜绿假单胞菌，宿主免疫防御的各个组成部分以及感染的小分子调节剂的作用。

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