Abstract
Skin is the primary barrier against the external environment and develops a robust immune network for its surveillance. The origin of the resident immune cells of the skin has become a focus of interest over past a decade. Fate mapping studies have revealed that the macrophages home into the skin as early as E12.5 and are derived from the yolk sac and fetal liver. The resident γδT cells are born in the thymus and home to the skin by E16.5. Recent work from our lab has shown that the embryonic macrophages can actively remodel the extracellular matrix in skin suggesting that the skin immune system can be activated long before exposure to foreign antigens. In this chapter, we present a detailed protocol for isolating monocytes, macrophages, and epidermal dendritic T cell populations from embryonic skin.
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References
Nestle FO, Di Meglio P et al (2009) Skin immune sentinels in health and disease. Nat Rev Immunol 9(10):679–679
Heath WR, Carbone FR (2013) The skin-resident and migratory immune system in steady state and memory: innate lymphocytes, dendritic cells and T cells. Nat Immunol 14(10):978–985
Jiang X et al (2010) Embryonic trafficking of gammadelta T cells to skin is dependent on E/P selectin ligands and CCR4. Proc Natl Acad Sci U S A 107(16):7443–7448
Gurish MF, Austen KF (2012) Developmental origin and functional specialization of mast cell subsets. Immunity 37(1):25–33
Hoeffel G et al (2012) Adult Langerhans cells derive predominantly from embryonic fetal liver monocytes with a minor contribution of yolk sac-derived macrophages. J Exp Med 209(6):1167–1181
Castellana D et al (2014) Macrophages contribute to the cyclic activation of adult hair follicle stem cells. PLoS Biol 12(12):e1002002
Tay SS et al (2013) The skin-resident immune network. Curr Dermatol Rep 3:13–22
Jameson J et al (2002) A role for skin gammadelta T cells in wound repair. Science 296(5568):747–749
Kurbet AS et al (2016) Sterile inflammation enhances ECM degradation in integrin β1 KO embryonic skin. Cell Rep 16(12):3334–3347
Sharp LL, Jameson JM, Cauvi G, Havran WL (2005) Dendritic epidermal T cells regulate skin homeostasis through local production of insulin-like growth factor 1. Nat Immunol 6(1):73–79
Epelman S, Lavine KJ, Randolph GJ (2014) Origin and functions of tissue macrophages. Immunity 41(1):21–35
Acknowledgments
We would like to thank H. Krishnamurthy and the Central Imaging and Flow Facility (CIFF) at NCBS for the use of the confocal microscopes and FACs facility. Animal work was partially supported by the National Mouse Research Resource (NaMoR) grant (BT/PR5981/MED/31/181/2012; 2013-2016) from the DBT. We thank members of the Raghavan lab for feedback. The SR lab is funded through core funds from inStem supported by the Department of Biotechnology (DBT), DBT grant (BT/PR8655/AGR/36/759/2013), and DST-SERB grant (EMR/2016/003199).
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Kurbet, A.S., Raghavan, S. (2018). Isolating Immune Cells from Mouse Embryonic Skin. In: Turksen, K. (eds) Skin Stem Cells. Methods in Molecular Biology, vol 1879. Humana Press, New York, NY. https://doi.org/10.1007/7651_2018_148
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DOI: https://doi.org/10.1007/7651_2018_148
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