Abstract
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.
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Acknowledgments
We want to thank all the members of the Bart Lambrecht and Hamida Hammad laboratory for their contribution to the development and optimization of these protocols.
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De Leeuw, E., Bosteels, C., Lambrecht, B.N., Hammad, H. (2022). Isolation of Conventional Murine Lung Dendritic Cell Subsets. In: Gorska, M.M. (eds) Asthma. Methods in Molecular Biology, vol 2506. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2364-0_17
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DOI: https://doi.org/10.1007/978-1-0716-2364-0_17
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