Abstract
Pulmonary dendritic cells (DCs) are potent antigen-presenting cells that can activate both naïve and memory/effector T cells. However, very little is known of how movements and localization of DCs contribute to these events. To study this, we have developed new procedures that combine precision-cut lung slices with cell staining using fluorescently tagged antibodies to detect individual cell types. In this chapter, we describe these methods in detail and show how they can be used to study the localization of not only DCs but also other leukocytes of interest, as well as structural cells within the lung.
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References
Lambrecht BN, Hammad H (2009) Biology of lung dendritic cells at the origin of asthma. Immunity 31(3):412–424. https://doi.org/10.1016/j.immuni.2009.08.008
Steinman RM (2007) Lasker Basic Medical Research Award. Dendritic cells: versatile controllers of the immune system. Nat Med 13(10):1155–1159. https://doi.org/10.1038/nm1643
Jakubzick C, Bogunovic M, Bonito AJ, Kuan EL, Merad M, Randolph GJ (2008) Lymph-migrating, tissue-derived dendritic cells are minor constituents within steady-state lymph nodes. J Exp Med 205(12):2839–2850. https://doi.org/10.1084/jem.20081430
Julia V, Hessel EM, Malherbe L, Glaichenhaus N, O'Garra A, Coffman RL (2002) A restricted subset of dendritic cells captures airborne antigens and remains able to activate specific T cells long after antigen exposure. Immunity 16(2):271–283
Lyons-Cohen MR, Thomas SY, Cook DN, Nakano H (2017) Precision-cut mouse lung slices to visualize live pulmonary dendritic cells. J Vis Exp (122):e55465. https://doi.org/10.3791/55465
Acknowledgments
We thank Michael Sanderson, Jun Chen, and Keiko Nakano for their advice and help with preparing lung slices and Jeff Tucker and Erica Scappini for microscopic analysis. This work was supported by the Intramural Research Program of the National Institutes of Health and the National Institute of Environmental Health Sciences.
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Lyons-Cohen, M.R., Nakano, H., Thomas, S.Y., Cook, D.N. (2018). Imaging Precision-Cut Lung Slices to Visualize Leukocyte Localization and Trafficking. In: Reinhardt, R. (eds) Type 2 Immunity. Methods in Molecular Biology, vol 1799. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-7896-0_18
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DOI: https://doi.org/10.1007/978-1-4939-7896-0_18
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Publisher Name: Humana, New York, NY
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