Laboratory of Immunoregulation and Mucosal Immunology, VIB Center for Inflammation Research, Ghent, Belgium.
Department of Internal Medicine, Ghent University, Ghent, Belgium.
Methods Mol Biol. 2022;2506:237-255. doi: 10.1007/978-1-0716-2364-0_17.
In the lungs, immune cells make contact with different antigens every day. This requires an adequate immune response. Dendritic cells (DCs) form a dense network in the respiratory mucosa and continuously sample inhaled allergens. They play an important role in bridging innate and adaptive immunity. DCs are classically divided into plasmacytoid DCs (pDCs) and conventional DCs (cDCs). cDCs in the steady-state are further subdivided into cDC1s and cDC2s based on their ontogeny and distinct non-redundant functions. Recently, a hyperactivated state of cDC2s has been described that arises during inflammation, coined inflammatory cDC2s (inf-cDC2s) that phenotypically mimics monocyte-derived cells and has a hybrid cDC1/macrophage functional identity. This chapter describes different enrichment methods and a fluorescence-activated cell sorting protocol that in combination allow for discrimination and isolation of pure DC subsets from the murine lung. The chapter represents an up-to-date, universal framework that can be adapted to other tissues and species which is an added value in intra- and interspecies comparative research.
在肺部，免疫细胞每天都会与不同的抗原接触。这需要适当的免疫反应。树突状细胞（DCs）在呼吸道黏膜中形成密集的网络，并不断对吸入的过敏原进行采样。它们在桥接先天免疫和适应性免疫方面发挥着重要作用。DCs 通常分为浆细胞样 DC（pDCs）和常规 DC（cDCs）。稳态下的 cDC 进一步根据其发生和独特的非冗余功能分为 cDC1 和 cDC2。最近，描述了一种 cDC2 的过度激活状态，这种状态在炎症期间出现，被称为炎性 cDC2（inf-cDC2），其表型类似于单核细胞衍生的细胞，并具有混合的 cDC1/巨噬细胞功能特性。本章描述了不同的富集方法和荧光激活细胞分选（FACS）方案，这些方法结合使用可以从鼠肺中区分和分离出纯 DC 亚群。该章节代表了一个最新的、通用的框架，可以适应其他组织和物种，这在种内和种间比较研究中是一个附加值。
