The avian respiratory tract is a common entry route for many pathogens and an important delivery route for vaccination in the poultry industry. Immune responses in the avian lung have mostly been studied in vivo due to the lack of robust, relevant in vitro and ex vivo models mimicking the microenvironment. Precision-cut lung slices (PCLS) have the major advantages of maintaining the 3-dimensional architecture of the lung and includes heterogeneous cell populations. PCLS have been obtained from a number of mammalian species and from chicken embryos. However, as the embryonic lung is physiologically undifferentiated and immunologically immature, it is less suitable to examine complex host-pathogen interactions including antimicrobial responses. Here we prepared PCLS from immunologically mature chicken lungs, tested different culture conditions, and found that serum supplementation has a detrimental effect on the quality of PCLS. Viable cells in PCLS remained present for ≥ 40 days, as determined by viability assays and sustained motility of fluorescent mononuclear phagocytic cells. The PCLS were responsive to lipopolysaccharide stimulation, which induced the release of nitric oxide, IL-1β, type I interferons and IL-10. Mononuclear phagocytes within the tissue maintained phagocytic activity, with live cell imaging capturing interactions with latex beads and an avian pathogenic Escherichia coli strain. Finally, the PCLS were also shown to be permissive to infection with low pathogenic avian influenza viruses. Taken together, immunologically mature chicken PCLS provide a suitable model to simulate live organ responsiveness and cell dynamics, which can be readily exploited to examine host-pathogen interactions and inflammatory responses.

中文翻译：

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精密切割肺切片：一种检查鸡肺中宿主与病原体相互作用的新型多功能工具。

禽呼吸道是许多病原体的常见进入途径，也是家禽业疫苗接种的重要传递途径。由于缺乏模拟微环境的稳健、相关的体外和离体模型，禽肺的免疫反应主要在体内研究。精密切割肺切片 (PCLS) 的主要优点是维持肺部的 3 维结构并包含异质细胞群。PCLS 已从许多哺乳动物物种和鸡胚胎中获得。然而，由于胚胎肺在生理上未分化且免疫学上不成熟，因此不太适合检查复杂的宿主-病原体相互作用，包括抗菌反应。在这里，我们从免疫成熟的鸡肺中制备PCLS，测试了不同的培养条件，发现血清补充对PCLS的质量有不利影响。根据活力测定和荧光单核吞噬细胞的持续运动性确定，PCLS 中的活细胞保持存在 ≥ 40 天。PCLS 对脂多糖刺激有反应，脂多糖刺激诱导一氧化氮、IL-1β、I 型干扰素和 IL-10 的释放。组织内的单核吞噬细胞保持吞噬活性，活细胞成像捕捉到与乳胶珠和禽类致病性大肠杆菌菌株的相互作用。最后，PCLS 还被证明允许感染低致病性禽流感病毒。总而言之，免疫成熟的鸡 PCLS 提供了一个合适的模型来模拟活器官反应性和细胞动力学，可以很容易地利用它来检查宿主-病原体相互作用和炎症反应。