Madenspacher Jennifer H, Fessler Michael B
Immunity, Inflammation and Disease Laboratory, National Institute of Environmental Health Sciences (NIEHS), National Institutes of Health.
Immunity, Inflammation and Disease Laboratory, National Institute of Environmental Health Sciences (NIEHS), National Institutes of Health;
J Vis Exp. 2016 Sep 28(115):54508. doi: 10.3791/54508.
Although community-acquired pneumonia remains a major public health problem, murine models of bacterial pneumonia have recently facilitated significant preclinical advances in our understanding of the underlying cellular and molecular pathogenesis. In vivo mouse models capture the integrated physiology and resilience of the host defense response in a manner not revealed by alternative, simplified ex vivo approaches. Several methods have been described in the literature for intrapulmonary inoculation of bacteria in mice, including aerosolization, intranasal delivery, peroral endotracheal cannulation under 'blind' and visualized conditions, and transcutaneous endotracheal cannulation. All methods have relative merits and limitations. Herein, we describe in detail a non-invasive, technically non-intensive, inexpensive, and rapid method for intratracheal delivery of bacteria that involves aspiration (i.e., inhalation) by the mouse of an infectious inoculum pipetted into the oropharynx while under general anesthesia. This method can be used for pulmonary delivery of a wide variety of non-caustic biological and chemical agents, and is relatively easy to learn, even for laboratories with minimal prior experience with pulmonary procedures. In addition to describing the aspiration pneumonia method, we also provide step-by-step procedures for assaying the subsequent in vivo pulmonary innate immune response of the mouse, in particular, methods for quantifying bacterial clearance and the cellular immune response of the infected airway. This integrated and simple approach to pneumonia assessment allows for rapid and robust evaluation of the effect of genetic and environmental manipulations upon pulmonary innate immunity.
尽管社区获得性肺炎仍然是一个重大的公共卫生问题，但细菌性肺炎的小鼠模型最近在我们对潜在细胞和分子发病机制的理解方面推动了显著的临床前进展。体内小鼠模型以一种替代的、简化的体外方法所未揭示的方式捕捉宿主防御反应的综合生理学和恢复力。文献中已经描述了几种在小鼠肺内接种细菌的方法，包括雾化、鼻内给药、在“盲视”和可视化条件下经口气管插管以及经皮气管插管。所有方法都有其相对的优点和局限性。在此，我们详细描述一种用于气管内输送细菌的非侵入性、技术要求不高、成本低廉且快速的方法，该方法涉及在全身麻醉下，小鼠吸入移液管滴入口咽部的感染性接种物（即吸入）。这种方法可用于多种无腐蚀性生物和化学制剂的肺部给药，并且相对容易掌握，即使对于之前在肺部操作方面经验极少的实验室也是如此。除了描述吸入性肺炎方法外，我们还提供了检测小鼠随后体内肺部固有免疫反应的分步程序，特别是量化细菌清除和感染气道细胞免疫反应的方法。这种综合且简单的肺炎评估方法能够快速且有力地评估基因和环境操作对肺部固有免疫的影响。
